論文 - 柴田　秀樹

Calmodulin enhances mTORC1 signaling by preventing TSC2-Rheb binding Open Access

Amemiya, Y; Ioi, Y; Araki, M; Kontani, K; Maki, M; Shibata, H; Takahara, T

JOURNAL OF BIOLOGICAL CHEMISTRY   301 巻 ( 2 ) 頁： 108122 - 108122   2025年2月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Biological Chemistry  

The mechanistic target of rapamycin complex 1 (mTORC1) functions as a master regulator of cell growth and proliferation. We previously demonstrated that intracellular calcium ion (Ca2+) concentration modulates the mTORC1 pathway via binding of the Ca2+ sensor protein calmodulin (CaM) to tuberous sclerosis complex 2 (TSC2), a critical negative regulator of mTORC1. However, the precise molecular mechanism by which Ca2+/CaM modulates mTORC1 activity remains unclear. Here, we performed a binding assay based on nano-luciferase reconstitution, a method for detecting weak interactions between TSC2 and its target, Ras homolog enriched in the brain (Rheb), an activator of mTORC1. CaM inhibited the binding of TSC2 to Rheb in a Ca2+-dependent manner. Live-cell imaging analysis indicated increased interaction between the CaM-binding region of TSC2 and CaM in response to elevated intracellular Ca2+ levels. Furthermore, treatment with carbachol, an acetylcholine analog, elevated intracellular Ca2+ levels and activated mTORC1. Notably, carbachol-induced activation of mTORC1 was inhibited by CaM inhibitors, corroborating the role of Ca2+/CaM in promoting the mTORC1 pathway. Consistent with the effect of Ca2+/CaM on the TSC2-Rheb interaction, increased intracellular Ca2+ concentration promoted the dissociation of TSC2 from lysosomes without affecting Akt-dependent phosphorylation of TSC2, suggesting that the regulatory mechanism of TSC2 by Ca2+/CaM is distinct from the previously established action mechanism of TSC2. Collectively, our findings offer mechanistic insights into TSC2-Rheb regulation mediated by Ca2+/CaM, which links Ca2+ signaling to mTORC1 activation.

DOI： 10.1016/j.jbc.2024.108122

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Cytoprotective Role of Autophagy in CDIP1 Expression-Induced Apoptosis in MCF-7 Breast Cancer Cells 査読有り 国際誌 Open Access

Inukai, R; Mori, K; Maki, M; Takahara, T; Shibata, H

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   25 巻 ( 12 )   2024年6月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：International Journal of Molecular Sciences  

Cell death-inducing p53-target protein 1 (CDIP1) is a proapoptotic protein that is normally expressed at low levels and is upregulated by genotoxic and endoplasmic reticulum stresses. CDIP1 has been reported to be localized to endosomes and to interact with several proteins, including B-cell receptor-associated protein 31 (BAP31) and apoptosis-linked gene 2 (ALG-2). However, the cellular and molecular mechanisms underlying CDIP1 expression-induced apoptosis remain unclear. In this study, we first demonstrated that CDIP1 was upregulated after treatment with the anticancer drug adriamycin in human breast cancer MCF-7 cells but was degraded rapidly in the lysosomal pathway. We also demonstrated that treatment with the cyclin-dependent kinase 5 (CDK5) inhibitor roscovitine led to an increase in the electrophoretic mobility of CDIP1. In addition, a phosphomimetic mutation at Ser-32 in CDIP1 resulted in an increase in CDIP1 expression-induced apoptosis. We also found that CDIP1 expression led to the induction of autophagy prior to apoptosis. Treatment of cells expressing CDIP1 with SAR405, an inhibitor of the class III phosphatidylinositol 3-kinase VPS34, caused a reduction in autophagy and promoted apoptosis. Therefore, autophagy is thought to be a defense mechanism against CDIP1 expression-induced apoptosis.

DOI： 10.3390/ijms25126520

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New Insights into the Regulation of mTOR Signaling via Ca<SUP>2+</SUP>-Binding Proteins 査読有り 国際誌 Open Access

Amemiya, Y; Maki, M; Shibata, H; Takahara, T

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   24 巻 ( 4 )   2023年2月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：International Journal of Molecular Sciences  

Environmental factors are important regulators of cell growth and proliferation. Mechanistic target of rapamycin (mTOR) is a central kinase that maintains cellular homeostasis in response to a variety of extracellular and intracellular inputs. Dysregulation of mTOR signaling is associated with many diseases, including diabetes and cancer. Calcium ion (Ca2+) is important as a second messenger in various biological processes, and its intracellular concentration is tightly regulated. Although the involvement of Ca2+ mobilization in mTOR signaling has been reported, the detailed molecular mechanisms by which mTOR signaling is regulated are not fully understood. The link between Ca2+ homeostasis and mTOR activation in pathological hypertrophy has heightened the importance in understanding Ca2+-regulated mTOR signaling as a key mechanism of mTOR regulation. In this review, we introduce recent findings on the molecular mechanisms of regulation of mTOR signaling by Ca2+-binding proteins, particularly calmodulin (CaM).

DOI： 10.3390/ijms24043923

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Hepatitis C Virus-Induced ROS/JNK Signaling Pathway Activates the E3 Ubiquitin Ligase Itch to Promote the Release of HCV Particles via Polyubiquitylation of VPS4A 査読有り 国際誌 Open Access

Deng, L; Liang, YJ; Ariffianto, A; Matsui, C; Abe, T; Muramatsu, M; Wakita, T; Maki, M; Shibata, H; Shoji, I

JOURNAL OF VIROLOGY   96 巻 ( 6 ) 頁： e0181121   2022年3月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Virology  

We previously reported that hepatitis C virus (HCV) infection activates the reactive oxygen species (ROS)/c-Jun N-terminal kinase (JNK) signaling pathway. However, the roles of ROS/JNK activation in the HCV life cycle remain unclear. We sought to identify a novel role of the ROS/JNK signaling pathway in the HCV life cycle. Immunoblot analysis revealed that HCV-induced ROS/JNK activation promoted phosphorylation of Itch, a HECT-type E3 ubiquitin ligase, leading to activation of Itch. The small interfering RNA (siRNA) knockdown of Itch significantly reduced the extracellular HCV infectivity titers, HCV RNA, and HCV core protein without affecting intracellular HCV infectivity titers, HCV RNA, and HCV proteins, suggesting that Itch is involved in the release of HCV particles. HCV-mediated JNK/Itch activation specifically promoted polyubiquitylation of an AAA-type ATPase, VPS4A, but not VPS4B, required to form multivesicular bodies. Site-directed mutagenesis revealed that two lysine residues (K23 and K121) on VPS4A were important for VPS4A polyubiquitylation. The siRNA knockdown of VPS4A, but not VPS4B, significantly reduced extracellular HCV infectivity titers. Coimmunoprecipitation analysis revealed that HCV infection specifically enhanced the interaction between CHMP1B, a subunit of endosomal sorting complexes required for transport (ESCRT)-III complex, and VPS4A, but not VPS4B, whereas VPS4A K23R/K121R greatly reduced the interaction with CHMP1B. HCV infection significantly increased ATPase activity of VPS4A, but not VPS4A K23R/K121R or VPS4B, suggesting that HCV-mediated polyubiquitylation of VPS4A contributes to activation of VPS4A. Taken together, we propose that the HCV-induced ROS/JNK/Itch signaling pathway promotes VPS4A polyubiquitylation, leading to enhanced VPS4ACHMP1B interaction and promotion of VPS4A ATPase activity, thereby promoting the release of HCV particles.

DOI： 10.1128/jvi.01811-21

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Amino Acid-Mediated Intracellular Ca<SUP>2+</SUP> Rise Modulates mTORC1 by Regulating the TSC2-Rheb Axis through Ca<SUP>2+</SUP>/Calmodulin 査読有り 国際誌 Open Access

Amemiya, Y; Nakamura, N; Ikeda, N; Sugiyama, R; Ishii, C; Maki, M; Shibata, H; Takahara, T

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   22 巻 ( 13 )   2021年7月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：International Journal of Molecular Sciences  

Mechanistic target of rapamycin complex 1 (mTORC1) is a master growth regulator by controlling protein synthesis and autophagy in response to environmental cues. Amino acids, especially leucine and arginine, are known to be important activators of mTORC1 and to promote lysosomal translocation of mTORC1, where mTORC1 is thought to make contact with its activator Rheb GTPase. Although amino acids are believed to exclusively regulate lysosomal translocation of mTORC1 by Rag GTPases, how amino acids increase mTORC1 activity besides regulation of mTORC1 subcellular localization remains largely unclear. Here we report that amino acids also converge on regulation of the TSC2-Rheb GTPase axis via Ca2+ /calmodulin (CaM). We showed that the amino acid-mediated increase of intracellular Ca2+ is important for mTORC1 activation and thereby contributes to the promotion of nascent protein synthesis. We found that Ca2+ /CaM interacted with TSC2 at its GTPase activating protein (GAP) domain and that a CaM inhibitor reduced binding of CaM with TSC2. The inhibitory effect of a CaM inhibitor on mTORC1 activity was prevented by loss of TSC2 or by an active mutant of Rheb GTPase, suggesting that a CaM inhibitor acts through the TSC2-Rheb axis to inhibit mTORC1 activity. Taken together, in response to amino acids, Ca2+ /CaM-mediated regulation of the TSC2-Rheb axis contributes to proper mTORC1 activation, in addition to the well-known lysosomal translocation of mTORC1 by Rag GTPases.

DOI： 10.3390/ijms22136897

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The Novel ALG-2 Target Protein CDIP1 Promotes Cell Death by Interacting with ESCRT-I and VAPA/B 査読有り Open Access

Inukai, R; Mori, K; Kuwata, K; Suzuki, C; Maki, M; Takahara, T; Shibata, H

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   22 巻 ( 3 ) 頁： 1 - 25   2021年2月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：International Journal of Molecular Sciences  

Apoptosis-linked gene 2 (ALG-2, also known as PDCD6) is a member of the penta-EFhand (PEF) family of Ca2+-binding proteins. The murine gene encoding ALG-2 was originally reported to be an essential gene for apoptosis. However, the role of ALG-2 in cell death pathways has remained elusive. In the present study, we found that cell death-inducing p53 target protein 1 (CDIP1), a pro-apoptotic protein, interacts with ALG-2 in a Ca2+-dependent manner. Co-immunoprecipitation analysis of GFP-fused CDIP1 (GFP-CDIP1) revealed that GFP-CDIP1 associates with tumor susceptibility gene 101 (TSG101), a known target of ALG-2 and a subunit of endosomal sorting complex required for transport-I (ESCRT-I). ESCRT-I is a heterotetrameric complex composed of TSG101, VPS28, VPS37 and MVB12/UBAP1. Of diverse ESCRT-I species originating from four VPS37 isoforms (A, B, C, and D), CDIP1 preferentially associates with ESCRT-I containing VPS37B or VPS37C in part through the adaptor function of ALG-2. Overexpression of GFP-CDIP1 in HEK293 cells caused caspase-3/7-mediated cell death. In addition, the cell death was enhanced by co-expression of ALG-2 and ESCRT-I, indicating that ALG-2 likely promotes CDIP1-induced cell death by promoting the association between CDIP1 and ESCRT-I. We also found that CDIP1 binds to vesicle-associated membrane protein-associated protein (VAP)A and VAPB through the two phenylalanines in an acidic tract (FFAT)-like motif in the C-terminal region of CDIP1, mutations of which resulted in reduction of CDIP1-induced cell death. Therefore, our findings suggest that different expression levels of ALG-2, ESCRT-I subunits, VAPA and VAPB may have an impact on sensitivity of anticancer drugs associated with CDIP1 expression.

DOI： 10.3390/ijms22031175

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The Penta-EF-Hand ALG-2 Protein Interacts with the Cytosolic Domain of the SOCE Regulator SARAF and Interferes with Ubiquitination 査読有り 国際誌 Open Access

Zhang, W; Muramatsu, A; Matsuo, R; Teranishi, N; Kahara, Y; Takahara, T; Shibata, H; Maki, M

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   21 巻 ( 17 ) 頁： 1 - 21   2020年9月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：International Journal of Molecular Sciences  

ALG-2 is a penta-EF-hand Ca2+-binding protein and interacts with a variety of proteins in mammalian cells. In order to find new ALG-2-binding partners, we searched a human protein database and retrieved sequences containing the previously identified ALG-2-binding motif type 2 (ABM-2). After selecting 12 high-scored sequences, we expressed partial or full-length GFP-fused proteins in HEK293 cells and performed a semi-quantitative in vitro binding assay. SARAF, a negative regulator of store-operated Ca2+ entry (SOCE), showed the strongest binding activity. Biochemical analysis of Strep-tagged and GFP-fused SARAF proteins revealed ubiquitination that proceeded during pulldown assays under certain buffer conditions. Overexpression of ALG-2 interfered with ubiquitination of wild-type SARAF but not ubiquitination of the F228S mutant that had impaired ALG-2-binding activity. The SARAF cytosolic domain (CytD) contains two PPXY motifs targeted by the WW domains of NEDD4 family E3 ubiquitin ligases. The PPXY motif proximal to the ABM-2 sequence was found to be more important for both in-cell ubiquitination and post-cell lysis ubiquitination. A ubiquitination-defective mutant of SARAF with Lys-to-Arg substitutions in the CytD showed a slower degradation rate by half-life analysis. ALG-2 promoted Ca2+-dependent CytD-to-CytD interactions of SARAF. The ALG-2 dimer may modulate the stability of SARAF by sterically blocking ubiquitination and by bridging SARAF molecules at the CytDs.

DOI： 10.3390/ijms21176315

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Amino acid-dependent control of mTORC1 signaling: a variety of regulatory modes 招待有り 査読有り 国際誌 Open Access

Takahara, T; Amemiya, Y; Sugiyama, R; Maki, M; Shibata, H

JOURNAL OF BIOMEDICAL SCIENCE   27 巻 ( 1 ) 頁： 87 - 87   2020年8月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Biomedical Science  

The mechanistic target of rapamycin complex 1 (mTORC1) is an essential regulator of cell growth and metabolism through the modulation of protein and lipid synthesis, lysosome biogenesis, and autophagy. The activity of mTORC1 is dynamically regulated by several environmental cues, including amino acid availability, growth factors, energy levels, and stresses, to coordinate cellular status with environmental conditions. Dysregulation of mTORC1 activity is closely associated with various diseases, including diabetes, cancer, and neurodegenerative disorders. The discovery of Rag GTPases has greatly expanded our understanding of the regulation of mTORC1 activity by amino acids, especially leucine and arginine. In addition to Rag GTPases, other factors that also contribute to the modulation of mTORC1 activity have been identified. In this review, we discuss the mechanisms of regulation of mTORC1 activity by particular amino acids.

DOI： 10.1186/s12929-020-00679-2

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ATeam technology for detecting early signs of viral cytopathic effect Open Access

Doysabas, KCC; Oba, M; Ishibashi, T; Shibata, H; Takemae, H; Shimoda, H; Tarigan, R; Mizutani, T; Iida, A; Hondo, E

JOURNAL OF VETERINARY MEDICAL SCIENCE   82 巻 ( 3 ) 頁： 387 - 393   2020年3月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：公益社団法人 日本獣医学会  

Adenosine 5’-triphosphate (ATP), the major energy currency of the cell, is involved in many cellular processes, including the viral life cycle, and can be used as an indicator of early signs of cytopathic effect (CPE). In this study, we demonstrated that CPE can be analyzed using an FRET-based ATP probe named ATP indicator based on Epsilon subunit for Analytical Measurements (ATeam). The results revealed that as early as 3 hr, the virus infected cells showed a significantly different Venus/cyan fluorescent protein (CFP) ratio compared to the mock-infected cells. The ATeam technology is therefore useful to determine the early signs of ATP-based CPE as early as 3 hr without morphology-based CPE by light microscopy, and enables high throughput determination of the presence of microorganisms in neglected samples stored in laboratories.

DOI： 10.1292/jvms.20-0021

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SH3YL1 cooperates with ESCRT-I in the sorting and degradation of the EGF receptor 査読有り 国際誌 Open Access

Hasegawa, J; Jebri, I; Yamamoto, H; Tsujita, K; Tokuda, E; Shibata, H; Maki, M; Itoh, T

JOURNAL OF CELL SCIENCE   132 巻 ( 19 )   2019年10月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Cell Science  

Ubiquitinated membrane proteins such as epidermal growth factor receptor (EGFR) are delivered to early endosomes and then sorted to lysosomes via multivesicular bodies (MVBs) for degradation. The regulatory mechanismunderlying formation of intralumenal vesicles en route to generation of MVBs is not fully understood. In this study, we found that SH3YL1, a phosphoinositide-binding protein, had a vesicular localization pattern overlapping with internalized EGF in endosomes in the degradative pathway. Deficiency of SH3YL1 prevents EGF trafficking from early to late endosomes and inhibits degradation of EGFR. Moreover, we show that SH3YL1 mediates EGFR sorting intoMVBs in a manner dependent on its C-terminal SH3 domain, which is necessary for the interaction with an ESCRT-I component, Vps37B. Taken together, our observations reveal an indispensable role of SH3YL1 in MVB sorting and EGFR degradation mediated by ESCRT complexes.

DOI： 10.1242/jcs.229179

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カルシウム依存的相互作用因子から探るpenta-EF-handファミリーの機能 査読有り

牧 正敏, 高原 照直, 柴田 秀樹

生化学   91 巻 ( 2 ) 頁： 191 - 209   2019年4月

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記述言語：日本語   出版者・発行元：公益社団法人日本生化学会  

典型的カルパイン分子中に見いだされたpenta-EF-hand（PEF）ドメインは，EF-handを連続して五つ持ち，C末端のEF5どうしが一対になって二量体を形成する．PEFファミリーには，sorcinやALG-2のように触媒ドメインを持たないメンバーが存在し，さまざまなタンパク質と相互作用してその機能を発揮する．動物細胞においてALG-2二量体はカルシウム依存性分子アダプターとして働き，異なる分子どうしの連結や標的複合体を安定化をする．ESCRTシステムにおける初期補助因子としての役割や，小胞体–ゴルジ体間における小胞輸送調節が注目されている．一方，PEFドメインを持たない非典型的カルパインであるcalpain-7は，N末端にMITドメインを持ち，ESCRT因子と相互作用して活性化され，エンドソーム・リソソーム経路で働いている．

DOI： 10.14952/seikagaku.2019.910191

CiNii Research

カルシウム依存的相互作用因子から探るpenta-EF-handファミリーの機能

牧 正敏, 高原 照直, 柴田 秀樹

生化学   91 巻 ( 2 ) 頁： 191 - 209   2019年4月

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記述言語：日本語   出版者・発行元：日本生化学会  

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その他リンク： http://search.jamas.or.jp/link/ui/2019233312

Adaptor functions of the Ca<SUP>2+</SUP>-binding protein ALG-2 in protein transport from the endoplasmic reticulum 招待有り 査読有り 国際誌 Open Access

Shibata, H

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   83 巻 ( 1 ) 頁： 20 - 32   2019年

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Bioscience, Biotechnology and Biochemistry  

Apoptosis-linked gene 2 (ALG-2) is a Ca2+-binding protein with five repetitive EF-hand motifs, named penta-EF-hand (PEF) domain. It interacts with various target proteins and functions as a Ca2+-dependent adaptor in diverse cellular activities. In the cytoplasm, ALG-2 is predominantly localized to a specialized region of the endoplasmic reticulum (ER), called the ER exit site (ERES), through its interaction with Sec31A. Sec31A is an outer coat protein of coat protein complex II (COPII) and is recruited from the cytosol to the ERES to form COPII-coated transport vesicles. I will overview current knowledge of the physiological significance of ALG-2 in regulating ERES localization of Sec31A and the following adaptor functions of ALG-2, including bridging Sec31A and annexin A11 to stabilize Sec31A at the ERES, polymerizing the Trk-fused gene (TFG) product, and linking MAPK1-interacting and spindle stabilizing (MISS)like (MISSL) and microtubule-associated protein 1B (MAP1B) to promote anterograde transport from the ER.

DOI： 10.1080/09168451.2018.1525274

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High Sensitive Quantitative Binding Assays Using a Nanoluciferase-Fused Probe for Analysis of ALG-2-Interacting Proteins 査読有り 国際誌

Zhang, W; Matsuo, R; Takahara, T; Shibata, H; Maki, M

CALCIUM-BINDING PROTEINS OF THE EF-HAND SUPERFAMILY   1929 巻   頁： 501 - 516   2019年

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Methods in Molecular Biology  

Many non-catalytic cellular proteins exert biological functions by formation of stable or transient complexes with other proteins. Analysis of the signal-induced physical interactions is important to understand their physiological roles in cells. Here we describe a biochemical method for assessing the binding of ALG-2 (gene name, PDCD6) to its target proteins that are immunoprecipitated from cell lysates. Application of nanoluciferase (Nluc)-fused ALG-2 enables a rapid quantitative evaluation of Ca 2+ -dependent interactions of target proteins with ALG-2 in vitro binding assays.

DOI： 10.1007/978-1-4939-9030-6_31

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Cellular Ca<SUP>2+</SUP>-Responding Nanoluciferase Reporter Gene System Directed by Tandemly Repeated Pseudo-palindromic NFAT-Response Elements 査読有り 国際誌

Zhang, W; Takahara, T; Achiha, T; Shibata, H; Maki, M

CALCIUM-BINDING PROTEINS OF THE EF-HAND SUPERFAMILY   1929 巻   頁： 95 - 109   2019年

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Methods in Molecular Biology  

Luciferase reporter gene systems based on the NFAT-response element (RE) have been used to monitor intracellular Ca 2+ elevation. However, Ca 2+ mobilization agent (e.g., ionomycin) alone is not adequate to activate the currently often employed reporter gene that contains the NFAT-RE found in the IL2 promoter. In addition to activation of NFAT through the Ca 2+ -calmodulin/calcineurin pathway, activation of AP-1 as a partner transcription factor is essential for the IL2-based NFAT-RE system. Here, we describe a detailed method for the recently developed new reporter gene system containing the NFAT-RE from the IL8 promoter. This system enables us to monitor endpoint effects of Ca 2+ -mobilizing agonists independent of AP-1 activation.

DOI： 10.1007/978-1-4939-9030-6_7

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Topics from special edition 小胞体からの分子輸送におけるALG-2の役割

柴田 秀樹, 高原 照直, 京 卓志, 牧 正敏

細胞   50 巻 ( 8 ) 頁： 441 - 445   2018年7月

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記述言語：日本語   出版者・発行元：ニューサイエンス社  

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その他リンク： http://search.jamas.or.jp/link/ui/2018298680

A microtubule-associated protein MAP1B binds to and regulates localization of a calcium-binding protein ALG-2 査読有り 国際誌

Takahara, T; Arai, Y; Kono, Y; Shibata, H; Maki, M

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   497 巻 ( 2 ) 頁： 492 - 498   2018年3月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Biochemical and Biophysical Research Communications  

MAP1B (microtubule-associated protein 1B) binds to microtubules and regulates microtubule dynamics. Previously, we showed calcium-dependent interaction between MAP1B and a calcium-binding protein ALG-2 (apoptosis-linked gene 2), which is involved in regulation of the protein secretion pathway. Although ALG-2 generally binds to proteins through two consensus binding motifs such as ABM-1 and ABM-2, the absence of these motifs in MAP1B suggests a unique binding mode between MAP1B and ALG-2. Here, we identified the region of mouse MAP1B responsible for binding to ALG-2, and found point mutations that abrogated binding of MAP1B to ALG-2. Furthermore, interaction between MAP1B and ALG-2 selectively prevented ALG-2 from binding to proteins with ABM-2 such as Sec31A, suggesting competition between MAP1B and ABM-2-containing proteins for binding to ALG-2. Consistently, in MAP1B knockout cells, co-localization of ALG-2 with Sec31A was increased. Moreover, overexpression of wild-type MAP1B, but not the MAP1B mutant defective in ALG-2 binding, altered localizations of ALG-2 and Sec31A into dispersed distributions, suggesting that MAP1B regulates localizations of ALG-2 and Sec31A in the cells. Finally, we found two cancer-associated mutations of human MAP1B located near ALG-2 binding sites. The introduction of the corresponding mutations in mouse MAP1B dramatically reduced the binding ability to ALG-2. Thus, these results suggest that MAP1B plays a role in regulation of ALG-2 and Sec31A localizations, and that dysregulation of calcium-dependent binding of ALG-2 to MAP1B might influence pathological conditions such as cancers.

DOI： 10.1016/j.bbrc.2018.02.048

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Nanoluciferase Reporter Gene System Directed by Tandemly Repeated Pseudo-Palindromic NFAT-Response Elements Facilitates Analysis of Biological Endpoint Effects of Cellular Ca<SUP>2+</SUP> Mobilization 査読有り 国際誌 Open Access

Zhang, W; Takahara, T; Achiha, T; Shibata, H; Maki, M

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   19 巻 ( 2 )   2018年2月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：International Journal of Molecular Sciences  

NFAT is a cytoplasm-localized hyper-phosphorylated transcription factor that is activated through dephosphorylation by calcineurin, a Ca2+/calmodulin-dependent phosphatase. A non-palindromic NFAT-response element (RE) found in the IL2 promoter region has been commonly used for a Ca2+-response reporter gene system, but requirement of concomitant activation of AP-1 (Fos/Jun) often complicates the interpretation of obtained results. A new nanoluciferase (NanoLuc) reporter gene containing nine-tandem repeats of a pseudo-palindromic NFAT-RE located upstream of the IL8 promoter was designed to monitor Ca2+-induced transactivation activity of NFAT in human embryonic kidney (HEK) 293 cells by measuring luciferase activities of NanoLuc and co-expressed firefly luciferase for normalization. Ionomycin treatment enhanced the relative luciferase activity (RLA), which was suppressed by calcineurin inhibitors. HEK293 cells that stably express human STIM1 and Orai1, components of the store-operated calcium entry (SOCE) machinery, gave a much higher RLA by stimulation with thapsigargin, an inhibitor of sarcoplasmic/endoplamic reticulum Ca2+-ATPase (SERCA). HEK293 cells deficient in a penta-EF-hand Ca2+-binding protein ALG-2 showed a higher RLA value than the parental cells by stimulation with an acetylcholine receptor agonist carbachol. The novel reporter gene system is found to be useful for applications to cell signaling research to monitor biological endpoint effects of cellular Ca2+ mobilization.

DOI： 10.3390/ijms19020605

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小胞体からの分子輸送におけるALG-2の役割

柴田 秀樹

月刊「細胞」   50 巻   頁： 441 - 445   2018年

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CiNii Research

The calcium-binding protein ALG-2 regulates protein secretion and trafficking via interactions with MISSL and MAP1B proteins. 査読有り

Terunao Takahara, Kuniko Inoue, Yumika Arai, Keiko Kuwata, Hideki Shibata, Masatoshi Maki

Journal of Biological Chemistry   292 巻 ( 41 ) 頁： 17057-17072   2017年10月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1074/jbc.M117.800201.

The calcium-binding protein ALG-2 regulates protein secretion and trafficking via interactions with MISSL and MAP1B proteins 査読有り 国際誌 Open Access

Takahara, T; Inoue, K; Arai, Y; Kuwata, K; Shibata, H; Maki, M

JOURNAL OF BIOLOGICAL CHEMISTRY   292 巻 ( 41 ) 頁： 17057 - 17072   2017年10月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Biological Chemistry  

Mobilization of intracellular calcium is essential for a wide range of cellular processes, including signal transduction, apo-ptosis, and vesicular trafficking. Several lines of evidence have suggested that apoptosis-linked gene 2 (ALG-2, also known as PDCD6), a calcium-binding protein, acts as a calcium sensor linking calcium levels with efficient vesicular trafficking, especially at the endoplasmic reticulum (ER)-to-Golgi transport step. However, how ALG-2 regulates these processes remains largely unclear. Here, we report that MAPK1-interacting and spindle-stabilizing (MISS)-like (MISSL), a previously uncharacterized protein, interacts with ALG-2 in a calcium-dependent manner. Live-cell imaging revealed that upon a rise in intracellular calcium levels, GFP-tagged MISSL (GFP-MISSL) dynamically relocalizes in a punctate pattern and colocalizes with ALG-2. MISSL knockdown caused disorganization of the components of the ER exit site, the ER-Golgi intermediate compartment, and Golgi. Importantly, knockdown of either MISSL or ALG-2 attenuated the secretion of secreted alkaline phospha-tase (SEAP), a model secreted cargo protein, with similar reductions in secretion by single- and double-protein knockdowns, suggesting that MISSL and ALG-2 act in the same pathway to regulate the secretion process. Furthermore, ALG-2 or MISSL knockdown delayed ER-to-Golgi transport of procollagen type I. We also found that ALG-2 and MISSL interact with microtubule-associated protein 1B (MAP1B) and that MAP1B knockdown reverts the reduced secretion of SEAP caused by MISSL or ALG-2 depletion. These results suggest that a change in the intracellular calcium level plays a role in regulation of the secretory pathway via interaction of ALG-2 with MISSL and MAP1B.

DOI： 10.1074/jbc.M117.800201

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Mutations in the vesicular trafficking protein annexin A11 are associated with amyotrophic lateral sclerosis 査読有り Open Access

Smith, BN; Topp, SD; Fallini, C; Shibata, H; Chen, HJ; Troakes, C; King, A; Ticozzi, N; Kenna, KP; Soragia-Gkazi, A; Miller, JW; Sato, A; Dias, DM; Jeon, M; Vance, C; Wong, CH; de Majo, M; Kattuah, W; Mitchell, JC; Scotter, EL; Parkin, NW; Sapp, PC; Nolan, M; Nestor, PJ; Simpson, M; Weale, M; Lek, M; Baas, F; de Jong, JMV; ten Asbroek, ALMA; Redondo, AG; Esteban-Pérez, J; Tiloca, C; Verde, F; Duga, S; Leigh, N; Pall, H; Morrison, KE; Al-Chalabi, A; Shaw, PJ; Kirby, J; Turner, MR; Talbot, K; Hardiman, O; Glass, JD; de Belleroche, J; Maki, M; Moss, SE; Miller, C; Gellera, C; Ratti, A; Al-Sarraj, S; Brown, RH; Silani, V; Landers, JE; Shaw, CE

SCIENCE TRANSLATIONAL MEDICINE   9 巻 ( 388 ) 頁： eaad9157   2017年5月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Science Translational Medicine  

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder. We screened 751 familial ALS patient whole-exome sequences and identified six mutations including p.D40G in the ANXA11 gene in 13 individuals. The p.D40G mutation was absent from 70,000 control whole-exome sequences. This mutation segregated with disease in two kindreds and was present in another two unrelated cases (P = 0.0102), and all mutation carriers shared a common founder haplotype. Annexin A11-positive protein aggregates were abundant in spinal cord motor neurons and hippocampal neuronal axons in an ALS patient carrying the p.D40G mutation. Transfected human embryonic kidney cells expressing ANXA11 with the p.D40G mutation and other N-terminal mutations showed altered binding to calcyclin, and the p.R235Q mutant protein formed insoluble aggregates. We conclude that mutations in ANXA11 are associated with ALS and implicate defective intracellular protein trafficking in disease pathogenesis. 2017

DOI： 10.1126/scitranslmed.aad9157

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The calcium-binding protein ALG-2 promotes endoplasmic reticulum exit site localization and polymerization of Trk-fused gene (TFG) protein 査読有り 国際誌 Open Access

Kanadome, T; Shibata, H; Kuwata, K; Takahara, T; Maki, M

FEBS JOURNAL   284 巻 ( 1 ) 頁： 56 - 76   2017年1月

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：FEBS Journal  

Apoptosis-linked gene 2 (ALG-2), which is a gene product of PDCD6, is a 22-kDa Ca2+-binding protein. Accumulating evidence points to a role for ALG-2 as a Ca2+-responsive adaptor protein. On binding to Ca2+, ALG-2 undergoes a conformational change that facilitates its interaction with various proteins. It also forms a homodimer and heterodimer with peflin, a paralog of ALG-2. However, the differences in cellular roles for the ALG-2 homodimer and ALG-2/peflin heterodimer are unclear. In the present study, we found that Trk-fused gene (TFG) protein interacted with the ALG-2 homodimer. Immunostaining analysis revealed that TFG and ALG-2 partially overlapped at endoplasmic reticulum exit sites (ERES), a platform for COPII-mediated protein transport from the endoplasmic reticulum. Time-lapse live-cell imaging demonstrated that both green fluorescent protein-fused TFG and mCherry-fused ALG-2 are recruited to ERES after thapsigargin treatment, which raises intracellular Ca2+ levels. Furthermore, overexpression of ALG-2 induced the accumulation of TFG at ERES. TFG has an ALG-2-binding motif and deletion of the motif decreased TFG binding to ALG-2 and shortened its half-life at ERES, suggesting a critical role for ALG-2 in retaining TFG at ERES. We also demonstrated, by in vitro cross-linking assays, that ALG-2 promoted the polymerization of TFG in a Ca2+-dependent manner. Collectively, the results suggest that ALG-2 acts as a Ca2+-sensitive adaptor to concentrate and polymerize TFG at ERES, supporting a potential role for ALG-2 in COPII-dependent trafficking from the endoplasmic reticulum.

DOI： 10.1111/febs.13949

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Multifaceted Roles of ALG-2 in Ca²⁺-Regulated Membrane Trafficking. 査読有り

Masatoshi Maki, Terunao Takahara, Hideki Shibata

International Journal of Molecular Sciences   17 巻 ( 9 ) 頁： E1401   2016年8月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3390/ijms17091401.

Tubby-like protein superfamily member PLSCR3 functions as a negative regulator of adipogenesis in mouse 3T3-L1 preadipocytes by suppressing induction of late differentiation stage transcription factors. 査読有り

Akira Inokawa, Tatsutoshi Inuzuka, Terunao Takahara, Hideki Shibata, Masatoshi Maki

Bioscience Reports   36 巻 ( 1 ) 頁： e00287   2016年1月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1042/BSR20150215

A New Role for Annexin A11 in the Early Secretory Pathway via Stabilizing Sec31A Protein at the Endoplasmic Reticulum Exit Sites (ERES). 査読有り

Hideki Shibata, Takashi Kanadome, Hirofumi Sugiura, Takeru Yokoyama, Minami Yamamuro, Stephen E. Moss, Masatoshi Maki

The Journal of Biological Chemistry   290 巻 ( 8 ) 頁： 4981-4993   2015年2月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1074/jbc.M114.592089

Structural Analysis of the Complex between Penta-EF-Hand ALG-2 Protein and Sec31A Peptide Reveals a Novel Target Recognition Mechanism of ALG-2 査読有り

Takeshi Takahashi, Kyosuke Kojima, Wei Zhang, Kanae Sasaki, Masaru Ito, Hironori Suzuki, Masato Kawasaki, Soichi Wakatsuki, Terunao Takahara, Hideki Shibata, Masatoshi Maki

International Journal of Molecular Sciences   16 巻 ( 2 ) 頁： 3677-3699   2015年2月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3390/ijms16023677

Involvement of calpain-7 in epidermal growth factor receptor degradation via the endosomal sorting pathway. 査読有り

Yuki Maemoto, Yasuko Ono, Satomi Kiso, Hideki Shibata, Terunao Takahara, Hiroyuki Sorimachi, Masatoshi Maki

FEBS J.   281 巻 ( 16 ) 頁： 3642-3655   2014年7月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Nuclear ALG-2 protein interacts with Ca2+ homeostasis endoplasmic reticulum protein (CHERP) Ca2+-dependently and participates in regulation of alternative splicing of inositol trisphosphate receptor type 1 (IP3R1) pre-mRNA. 査読有り

Kanae Sasaki-Osugi, Chiaki Imoto, Terunao Takahara, Hideki Shibata, Masatoshi Maki

J. Biol. Chem.   288 巻 ( 46 ) 頁： 33361-33375   2013年9月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

VPS37 isoforms differentially modulate the ternary complex formation of ALIX, ALG-2, and ESCRT-I. 査読有り

Mayumi Okumura, Angela M. Katsuyama, Hideki Shibata, Masatoshi Maki

Biosci. Biotechnol. Biochem.   77 巻 ( 8 ) 頁： 1715-1721   2013年8月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Identification of phosphorylation sites in the C-terminal region of charged multivesicular body protein 1A (CHMP1A). 査読有り

Yuki Maemoto, Hideki Shibata, Masatoshi Maki

Biosci. Biotechnol. Biochem.   77 巻 ( 6 ) 頁： 1317-1319   2013年6月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Mammalian ESCRT-III-related protein IST1 has a distinctive Met-Pro repeat sequence that is essential for interaction with ALG-2 in the presence of Ca2+. 査読有り

Mayumi Okumura, Takeshi Takahashi, Hideki Shibata, Masatoshi Maki

Biosci. Biotechnol. Biochem.   77 巻 ( 5 ) 頁： 1049-1054   2013年5月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Analysis of limited proteolytic activity of calpain-7 using non-physiological substrates in mammalian cells. 査読有り

Yuki Maemoto, Satomi Kiso, Hideki Shibata, Masatoshi Maki

FEBS J.   280 巻 ( 11 ) 頁： 2594-2607   2013年4月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

ALG-2-interacting Tubby-like protein superfamily member PLSCR3 is secreted by an exosomal pathway and taken up by recipient cultured cells. 査読有り

Tatsutoshi Inuzuka, Akira Inokawa, Cen Chen, Kumiko Kizu, Hiroshi Narita, Hideki Shibata, Masatoshi Maki

Biosci. Rep.   33 巻 ( 2 ) 頁： e00026   2013年3月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Biochemical and immunological detection of physical interactions between penta-EF-hand protein ALG-2 and its binding partners. 査読有り

Kanae Osugi, Hideki Shibata, Masatoshi Maki

Methods Mol Biol.   963 巻   頁： 187-200   2012年11月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1007/978-1-62703-230-8_12.

Prediction of a New Ligand-Binding Site for Type 2 Motif based on the Crystal Structure of ALG-2 by Dry and Wet Approaches. 査読有り

Takeshi Takahashi, Hironori Suzuki, Tatsutoshi Inuzuka, Hideki Shibata, Masatoshi Maki

Int J Mol Sci.   13 巻 ( 6 ) 頁： 7535-7549   2012年6月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Identification of the P-body component PATL1 as a novel ALG-2-interacting protein by in silico and far-Western screening of proline-rich proteins. 査読有り

Kanae Osugi, Hironori Suzuki, Tomomi Nomura, Yasuo Ariumi, Hideki Shibata, Masatoshi Maki

J Biochem.   151 巻 ( 6 ) 頁： 657-666   2012年6月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Evolutionary and physical linkage between calpains and penta-EF-hand Ca2+-binding proteins. 査読有り

Masatoshi Maki, Yuki Maemoto, Yohei Osako, Hideki Shibata

FEBS J.   279 巻 ( 8 ) 頁： 1414-1421   2012年4月

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記述言語：英語  

DOI： 10.1111/j.1742-4658.2012.08560.x.

Calpain-7 binds to CHMP1B at its second α-helical region and forms a ternary complex with IST1. 査読有り

Yuki Maemoto, Yohei Osako, Emi Goto, Eri Nozawa, Hideki Shibata, Masatoshi Maki

J. Biochem.   150 巻 ( 4 ) 頁： 411-421   2011年10月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Structure and function of ALG-2, a penta-EF-hand calcium-dependent adaptor protein. 査読有り

Masatoshi Maki, Hironori Suzuki, Hideki Shibata

Sci China Life Sci.   54 巻 ( 8 ) 頁： 770-779   2011年8月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Autolytic activity of human calpain 7 is enhanced by ESCRT-III-related protein IST1 through MIT-MIM interaction. 査読有り

Yohei Osako, Yuki Maemoto, Ryohei Tanaka, Hironori Suzuki, Hideki Shibata, Masatoshi Maki

FEBS J.   277 巻 ( 21 ) 頁： 4412-4426   2010年11月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

*The ALG-2 binding site in Sec31A influences the retention kinetics of Sec31A at the endoplasmic reticulum exit sites as revealed by live-cell time-lapse imaging. 査読有り

Shibata H, Inuzuka T, Yoshida H, Sugiura H, Wada I, Maki M.

Biosci Biotechnol Biochem.   74 巻 ( 9 ) 頁： 1819-1826   2010年9月

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担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

ALG-2, a member of the penta-EF-hand protein family, interacts Ca²+-dependently with a COPII component, Sec31A. In this study, we first established HeLa cells stably expressing green fluorescent protein-fused ALG-2 (GFP-ALG-2) and red fluorescent protein-fused Sec31A (Sec31A-RFP). After inducing Ca²+-mobilization, the cytoplasmic distribution of GFP-ALG-2 changed from a diffuse to a punctate pattern, which extensively overlapped with the Sec31A-RFP-positive structures, indicating that ALG-2 is recruited to the endoplasmic reticulum exit sites (ERES) in living cells. Next, overlay experiments with biotin-labeled ALG-2 were done to dissect the ALG-2 binding site (ABS). They revealed that a sequence comprising amino acid residues 839-851 in the Pro-rich region was necessary and sufficient for direct binding to ALG-2. Finally, fluorescence recovery after photobleaching analysis indicated that the ABS deletion reduced the high-affinity population of Sec31A to the ERES, suggesting that the ABS is one of the key determinants of the retention kinetics of Sec31A at ERES.

Distinct functions of human MVB12A and MVB12B in the ESCRT-I dependent on their posttranslational modifications. 査読有り

Takumi Tsunematsu, Emiko Yamauchi, Hideki Shibata, Masatoshi Maki, Takeshi Ohta, Hiroaki Konishi

Biochem. Biophys. Res. Commun.   399 巻 ( 2 ) 頁： 232-237   2010年8月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Molecular basis for defect in Alix-binding by alternatively spliced isoform of ALG-2 (ALG-2DeltaGF122) and structural roles of F122 in target recognition. 査読有り

Tatsutoshi Inuzuka, Hironori Suzuki, Masato Kawasaki, Hideki Shibata, Soichi Wakatsuki, Masatoshi Maki

BMC Struct. Biol.   10 巻   2010年8月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： doi:10.1186/1472-6807-10-25

*Penta-EF-hand protein ALG-2 functions as a Ca2+-dependent adaptor that bridges Alix and TSG101. 査読有り

Okumura M, Ichioka F, Kobayashi R, Suzuki H, Yoshida H, Shibata H, Maki M.

Biochem Biophys Res Commun.   386 巻 ( 1 ) 頁： 237-241   2009年8月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The mechanism of Ca2+-dependent recognition of Alix by ALG-2: insights from X-ray crystal structures. 査読有り

Suzuki H, Kawasaki M, Inuzuka T, Okumura M, Kakiuchi T, Shibata H, Wakatsuki S, Maki M.

Biochem Soc Trans.   37 巻 ( Pt1 ) 頁： 190-194   2009年2月

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記述言語：英語  

Crystallization and X-ray diffraction analysis of N-terminally truncated human ALG-2. 査読有り

Hironori Suzuki, Masato Kawasaki, Tatsutoshi Inuzuka, Mayumi Okumura, Takeshi Kakiuchi, Hideki Shibata, Soichi Wakatsuki, Masatoshi Maki

Acta Crystallogr Sect F Struct Biol Cryst Commun.   64 巻 ( Pt11 ) 頁： 974-977   2008年11月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

*Structural basis for Ca2+-dependent formation of ALG-2/Alix peptide complex: Ca2+/EF3-driven arginine switch mechanism. 査読有り

Hironori Suzuki, Masato Kawasaki, Tatsutoshi Inuzuka, Mayumi Okumura, Takeshi Kakiuchi, Hideki Shibata, Soichi Wakatsuki, Masatoshi Maki

Structure   16 巻 ( 10 ) 頁： 1562-1573   2008年10月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

*Human calpain7/PalBH associates with a subset of ESCRT-III-related proteins in its N-terminal region and partly localizes to endocytic membrane compartments. 査読有り

Chiharu Yorikawa, Emi Takaya, Yohei Osako, Ryohei Tanaka, Yoshinori Terasawa, Takao Hamakubo, Yasuhiro Mochizuki, Hiroko Iwanari, Tatsuhiko Kodama, Tatsuya Maeda, Kiyotaka Hitomi, Hideki Shibata, Masatoshi Maki

J. Biochem.   143 巻 ( 6 ) 頁： 731-745   2008年6月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

*Identification of Alix-type and Non-Alix-type ALG-2-binding sites in human phospholipid scramblase 3: differential binding to an alternatively spliced isoform and amino acid-substituted mutants. 査読有り

Hideki Shibata, Hironori Suzuki, Takeshi Kakiuchi, Tatsutoshi Inuzuka, Haruna Yoshida, Takako Mizuno, Masatoshi Maki

J. Biol. Chem.   283 巻 ( 15 ) 頁： 9623-9632   2008年4月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Brox, a novel farnesylated Bro1 domain-containing protein that associates with charged multivesicular body protein 4 (CHMP4). 査読有り

Fumitaka Ichioka, Ryota Kobayashi, Keiichi Katoh, Hideki Shibata, Masatoshi Maki

FEBS J.   275 巻 ( 4 ) 頁： 682-692   2008年2月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Identification of substrates for transglutaminase in Physarum polycephalum, an acellular slime mold, upon cellular mechanical damage. 査読有り

Fumitaka Wada, Hiroki Hasegawa, Akio Nakamura, Yoshiaki Sugimura, Yoshiki Kawai, Narie Sasaki, Hideki Shibata, Masatoshi Maki, Kiyotaka Hitomi

FEBS J.   274 巻 ( 11 ) 頁： 2766-2777   2007年4月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

*ALG-2 directly binds Sec31A and localizes at endoplasmic reticulum exit sites in a Ca2+-dependent manner. 査読有り

Hideki Shibata, Hironori Suzuki, Haruna Yoshida, Masatoshi Maki

Biochem. Biophys. Res. Commun.   353 巻 ( 3 ) 頁： 756-763   2007年2月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

HD-PTP and Alix share some membrane-traffic related proteins that interact with their Bro1 domains or proline-rich regions. 査読有り

Fumitaka Ichioka, Emi Takaya, Hironori Suzuki, Sachiko Kajigaya, Vladimir L. Buchman, Hideki Shibata, Masatoshi Maki

Arch. Biochem. Biophys.   457 巻 ( 2 ) 頁： 142-149   2007年1月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The penta-EF-hand protein ALG-2 and its interacting proteins. 査読有り

Masatoshi Maki, Hideki Shibata

Calcium Binding Proteins   2 巻   頁： 4-10   2007年

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記述言語：英語  

CHMP7, a novel ESCRT-III-related protein, associates with CHMP4b and functions in the endosomal sorting pathway. 査読有り

Mio Horii, Hideki Shibata, Ryota Kobayashi, Keiichi Katoh, Chiharu Yorikawa, Jiro Yasuda, Masatoshi Maki

Biochem. J.   400 巻 ( 1 ) 頁： 23-32   2006年11月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Reevaluation of the Predicted Gene Structure of Dictyostelium Cyctatin A3 (cpiC) by Nucleotide Sequence Determination of its cDNA and its Phylogenetic Position in the Cyctatin Superfamily 査読有り

Medhat S. El-Halawany, Hideki Shibata, Kiyotaka Hitomi, Masatoshi Maki

Molecular Biology Reports   32 巻 ( 4 ) 頁： 257-264   2005年12月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The penta-EF-hand protein ALG-2 interacts directly with the ESCRT-I component TSG101, and Ca²⁺-dependently co-localizes to aberrant endosomes with dominant-negative AAA ATPase SKD1/Vps4B 査読有り

Keiichi Katoh, Hidenori Suzuki, Yoshinori Terasawa, Takako Mizuno, Jiro Yasuda, Hideki Shibata, Masatoshi Maki

Biochemical Journal   391 巻 ( Pt 3 ) 頁： 677-685   2005年11月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Identification of Rab GTPase-Activating Protein-Like Protein (RabGAPLP) as a Novel Alix/AIP1-Interacting Protein 査読有り

Fumitaka Ichioka, Mio Horii, Keiichi Katoh, Yoshinori Terasawa, Hideki Shibata, Masatoshi Maki

Bioscience, Biotechnology, and Biochemistry   69 巻 ( 4 ) 頁： 861-865   2005年4月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Human CHMP6, a myristoylated ESCRT-III protein, interacts directly with an ESCRT-II component EAP20 and regulates endosomal cargo sorting 査読有り

Chiharu Yorikawa, Hideki Shibata, Satoshi Waguri, Kazumi Hatta, Mio Horii, Keiichi Katoh, Toshihide Kobayashi, Yasuo Uchiyama, Masatoshi Maki

Biochemical Journal   387 巻 ( Pt 1 ) 頁： 17-26   2005年4月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Dictyostelium discoideum requires an Alix/AIP1 homolog, DdAlix, for morphogenesis in alkaline environments 査読有り

Susumu Ohkouchi, Hajime Saito, Fumika Aruga, Tatsuya Maeda, Hideki Shibata, Masatoshi Maki

FEBS Letters   579 巻 ( 7 ) 頁： 1745-1750   2005年3月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DdAlix, an Alix/AIP1 homolog in Dictyostelium discoideum, is required for multicellular development under low Ca²⁺ conditions 査読有り

Susumu Ohkouchi, Medhat S. El-Halawany, Fumika Aruga, Hideki Shibata, Kiyotaka Hitomi, Masatoshi Maki

Gene   337 巻   頁： 131-139   2004年8月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Identification of cysteine protease inhibitors that belong to cystatin family 1 in the cellular slime mold Dictyostelium discoideum 査読有り

Medhat S. El-Halawany, Susumu Ohkouchi, Hideki Shibata, Kiyotaka Hitomi, Masatoshi Maki

Biological Chemistry   385 巻 ( 6 ) 頁： 547-550   2004年6月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Protein kinase PKN1 associates with TRAF2 and is involved in TRAF2-NF-kappaB signaling pathway 査読有り

Yusuke Gotoh, Kumiko Oishi, Hideki Shibata, Akiko Yamagiwa, Takayuki Isagawa, Tamako Nishimura, Emiko Goyama, Mikiko Takahashi, Hideyuki Mukai, Yoshitaka Ono

Biochemical and Biophysical Research Communications   314 巻 ( 3 ) 頁： 688-694   2004年2月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The Penta-EF-Hand Protein ALG-2 Interacts with a Region Containing PxY Repeats in Alix/AIP1, Which Is Required for the Subcellular Punctate Distribution of the Amino-Terminal Truncation Form of Alix/AIP1 査読有り

Hideki Shibata, Keiko Yamada, Takako Mizuno, Chiharu Yorikawa, Hiroshi Takahashi, Hirokazu Satoh, Yasuyuki Kitaura, Masatoshi Maki

The Journal of Biochemistry   135 巻 ( 1 ) 頁： 117-128   2004年1月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

CHMP4b is a major binding partner of the ALG-2-interacting protein Alix among the three CHMP4 isoforms 査読有り

Keiichi Katoh, Hideki Shibata, Kazumi Hatta, Masatoshi Maki

Archives of Biochemistry and Biophysics   421 巻 ( 1 ) 頁： 159-165   2004年1月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The ALG-2-interacting Protein Alix Associates with CHMP4b, a Human Homologue of Yeast Snf7 That Is Involved in Multivesicular Body Sorting 査読有り

Keiichi Katoh, Hideki Shibata, Hidenori Suzuki, Atsuki Nara, Kazumi Ishidoh, Eiki Kominami, Tamotsu Yoshimori, Masatoshi Maki

The Journal of Biological Chemistry   278 巻 ( 40 ) 頁： 39104-39113   2003年10月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The penta-EF-hand domain of ALG-2 interacts with amino-terminal domains of both annexin VII and annexin XI in a Ca²⁺-dependent manner 査読有り

Hirokazu Satoh, Yoshimi Nakano, Hideki Shibata, Masatoshi Maki

Biochimica et Biophysica Acta   1600 巻 ( 1-2 ) 頁： 61-67   2002年11月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Structures, functions and molecular evolution of the penta-EF-hand Ca²⁺-binding proteins 査読有り

  1600 巻 ( 1-2 ) 頁： 51-60   2002年11月

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記述言語：英語  

ALG-2 Interacts with the Amino-Terminal Domain of Annexin XI in a Ca²⁺-Dependent Manner 査読有り

Hirokazu Satoh, Hideki Shibata, Yoshimi Nakano, Yasuyuki Kitaura, Masatoshi Maki

Biochemical and Biophysical Research Communications   291 巻 ( 5 ) 頁： 1166-1172   2002年3月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Both ALG-2 and Peflin, Penta-EF-hand (PEF) Proteins, Are Stabilized by Dimerization through Their Fifth EF-Hand Regions 査読有り

Yasuyuki Kitaura, Hirokazu Satoh, Hiroshi Takahashi, Hideki Shibata, Masatoshi Maki

Archives of Biochemistry and Biophysics   399 巻 ( 1 ) 頁： 12-18   2002年3月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

PKNbeta Interacts with the SH3 Domains of Graf and a Novel Graf Related Protein, Graf2, Which Are GTPase Activating Proteins for Rho Family 査読有り

Hideki Shibata, Kumiko Oishi, Akiko Yamagiwa, Mikiko Matsumoto, Hideyuki Mukai, Yoshitaka Ono

The Journal of Biochemistry   130 巻 ( 1 ) 頁： 23-31   2001年7月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Interaction of PKN with a neuron-specific basic helix-loop-helix transcription factor, NDRF/NeuroD2 査読有り

Hideki Shibata, Hisanobu Oda, Hideyuki Mukai, Kumiko Oishi, Kazuyo Misaki, Hiroaki Ohkubo, Yoshitaka Ono

Brain Reserch Molecular Brain Reserch   74 巻 ( 1-2 ) 頁： 126-134   1999年12月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Identification and characterization of PKNbeta, a novel isoform of protein kinase PKN: expression and arachidonic acid dependency are different from those of PKNalpha 査読有り

Kumiko Oishi, Hideyuki Mukai, Hideki Shibata, Mikiko Takahashi, Yoshitaka Ono

Biochemical and Biophysical Research Communications   261 巻 ( 3 ) 頁： 808-814   1999年8月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Characterization of a novel giant scaffolding protein, CG-NAP, that anchors multiple signaling enzymes to centrosome and the golgi apparatus 査読有り

Mikiko Takahashi, Hideki Shibata, Masaki Shimakawa, Masaaki Miyamoto, Hideyuki Mukai, Yoshitaka Ono

The Journal of Biological Chemistry   274 巻 ( 24 ) 頁： 17267-17274   1999年6月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Interaction of the small G protein RhoA with the C terminus of human phospholipase D1 査読有り

"Masakazu Yamazaki, Yue Zhang, Hiroshi Watanabe, Takeaki Yokozeki, Shigeo Ohno, Kozo Kaibuchi, Hideki Shibata, Hideyuki Mukai, Yoshitaka Ono, Michael A. Frohman, Yasunori Kanaho"

The Journal of Biological Chemistry   274 巻 ( 10 ) 頁： 6035-6038   1999年3月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

PKN interacts with a paraneoplastic cerebellar degeneration-associated antigen, which is a potential transcription factor 査読有り

Hiromi Takanaga, Hideyuki Mukai, Hideki Shibata, Masanao Toshimori, Yoshitaka Ono

Experimental Cell Research   241 巻 ( 2 ) 頁： 363-372   1998年6月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Domain-specific phosphorylation of vimentin and glial fibrillary acidic protein by PKN 査読有り

Kaori Matsuzawa, Hidetaka Kosako, Naoyuki Inagaki, Hideki Shibata, Hideyuki Mukai, Yoshitaka Ono, Mutsuki Amano, Kozo Kaibuchi, Yoshiharu Matsuura, Ichiro Azuma, Masaki Inagaki

Biochemical and Biophysical Research Communications   234 巻 ( 3 ) 頁： 621-625   1997年5月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Interaction of PKN with alpha-actinin 査読有り

Hideyuki Mukai, Masanao Toshimori, Hideki Shibata, Hiromi Takanaga, Michinori Kitagawa, Masako Miyahara, Masaki Shimakawa, Yoshitaka Ono

The Journal of Biological Chemistry   272 巻 ( 8 ) 頁： 4740-4746   1997年2月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Translocation of PKN from the cytosol to the nucleus induced by stresses 査読有り

Hideyuki Mukai, Masako Miyahara, Hiroko Sunakawa, Hideki Shibata, Masanao Toshimori, Michinori Kitagawa, Masaki Shimakawa, Hiromi Takanaga, Yoshitaka Ono

Proceedings of the National Academy of Sciences of the United States of America   93 巻 ( 19 ) 頁： 10195-10199   1996年9月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Characterization of the interaction between RhoA and the amino-terminal region of PKN. 査読有り

Hideki Shibata, Hideyuki Mukai,Yoshimasa Inagaki, Yoshimi Homma, Kazushi Kimura, Kozo Kaibuchi, Shuh Narumiya, Yoshitaka Ono

FEBS Letters   385 巻 ( 3 ) 頁： 221-224   1996年5月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

PKN associates and phosphorylates the head-rod domain of neurofilament protein 査読有り

Hideyuki Mukai, Masanao Toshimori, Hideki Shibata, Michinori Kitagawa, Masaki Shimakawa, Masako Miyahara, Hiroko Sunakawa, Yoshitaka Ono

The Journal of Biological Chemistry   271 巻 ( 16 ) 頁： 9816-9822   1996年4月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The role of the unique motifs in the amino-terminal region of PKN on its enzymatic activity 査読有り

Michinori Kitagawa, Hideki Shibata, Masanao Toshimori, Hideyuki Mukai, Yoshitaka Ono

Biochemical and Biophysical Research Communications   220 巻 ( 3 ) 頁： 963-968   1996年3月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Identification of Schizosaccharomyces pombe gene psk1⁺, encoding a novel putative serine/threonine protein kinase, whose mutation conferred resistance to phenylarsine oxide." 査読有り

Hideyuki Mukai, Masako Miyahara, Hiromi Takanaga, Michinori Kitagawa, Hideki Shibata, Masaki Shimakawa, Yoshitaka Ono

Gene   166 巻 ( 1 ) 頁： 155-159   1995年12月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Purification and characterization of a fatty acid-activated protein kinase (PKN) from rat testis 査読有り

Michinori Kitagawa, Hideyuki Mukai, Hideki Shibata, Yoshitaka Ono"

Biochemical Journal   310 巻 ( Pt 2 ) 頁： 657-664   1995年9月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Xenopus PKN: cloning and sequencing of the cDNA and identification of conserved domains 査読有り

Hideyuki Mukai, Kazuya Mori, Hiromi Takanaga, Michinori Kitagawa, Hideki Shibata, Masaki Shimakawa, Masako Miyahara, Yoshitaka Ono"

Biochimica et Biophysica Acta   1261 巻 ( 2 ) 頁： 296-300   1995年4月

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Activation of PKN, a novel 120-kDa protein kinase with leucine zipper- like sequences, by unsaturated fatty acids and by limited proteolysis 査読有り

Biochemical and Biophysical Research Communications   204 巻 ( 1 ) 頁： 348-356   1994年10月

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記述言語：英語   掲載種別：研究論文（学術雑誌）