記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.bbagen.2020.129679

Web of Science

PubMed

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1089/dna.2019.4700

Web of Science

PubMed

記述言語：日本語   掲載種別：研究論文（研究会，シンポジウム資料等）  

記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：公益社団法人 日本農芸化学会  

DOI： 10.1271/kagakutoseibutsu.57.205

CiNii Research

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1093/jb/mvy034

Web of Science

PubMed

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1002/cbf.3350

Web of Science

PubMed

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1111/gtc.12521

Web of Science

PubMed

記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：日本電気泳動学会  

<p>密度勾配超遠心分離とSDS-PAGEを用いて，マウス乳中に含まれる分泌膜小胞結合タンパク質，MFG-E8，の研究を行った．搾乳した新鮮なマウス乳に含まれる分泌膜小胞である乳脂肪球とエキソソームは密度勾配超遠心分離により，それぞれ低密度画分と高密度画分に分画され，SDS-PAGE免疫ブロット解析によりMFG-E8は主に乳脂肪球画分に検出された．このようなMFG-E8の分布は強制離乳により変化し，エキソソーム画分や可溶性タンパク質画分を含む幅広い密度範囲に分布した．乳脂肪球の蛍光免疫染色の結果などから，MFG-E8は分泌後間もない新鮮な乳脂肪球の表面には少なく時間経過とともに増加することが示された．MFG-E8の結合により乳脂肪球は貪食され易くなったことから，アポトーシス細胞と同様に，MFG-E8は古くなった乳脂肪球においてもeat-me signalとして機能することが示唆された．</p>

DOI： 10.2198/electroph.61.115

CiNii Research

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1080/09168451.2015.1116927

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1074/jbc.M114.598821

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1242/dev.121863

記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1007/s11010-014-2128-x

記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：日本語   掲載種別：研究論文（大学，研究機関等紀要）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： https://doi.org/10.1271/bbb.121011

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1093/jb/mvs154

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Milk fat globules (MFGs) secreted by lactating mammary gland are unique lipid surrounded by a phospholipid bi-layer. We report here post-weaning changes in milk fat globule EGF factor VIII (MFG-E8) and annexin V-accessible phosphatidylserine on the surface of MFGs. The MFG content in milk markedly decreased to about half within 2 days after forced weaning, despite a slight increase in milk protein content. Immunofluorescence-staining of MFGs using anti-MFG-E8 and annexin V indicated that MFG-E8 was present on some, but not all, MFGs before weaning, whereas most of MFGs were MFG-E8-positive and annexin V-negative after weaning. Free MFG-E8 with binding activity to phosphatidylserine was present abundantly in the post-weaning milk, and indeed exhibited binding to MFGs in pre-weaning milk. MFGs were taken up by HC11 mouse mammary epithelial cells in vitro, and those from post-weaning milk were remarkable for such cellular uptake. Moreover, the uptake of MFGs by the cells was inhibited by an anti-MFG-E8 antibody. Taken together, these findings suggest that MFG-E8 plays a critical role in regulation of MFG dynamics after weaning or during the suckling interval through the control of MFG-epithelial cell interaction in lactating mammary glands.

DOI： 10.1093/jb/mvs116

記述言語：英語   掲載種別：研究論文（学術雑誌）  

In lactating mammary glands, milk fat is secreted as fat globules surrounded by a cell plasma membrane containing characteristic membrane-associated proteins. Among these, butyrophilin has been shown to be specific and intrinsic to the fat globule membrane, whereas milk-fat globule EGF-factor VIII (MFG-E8) is uncertain. We characterized in the present study MFG-E8 in milk fat globules and in the culture medium of HC11 mammary epithelial cells. MFG-E8 was immunologically detected in the mammary tissues of both pregnant and lactating mice. Double-immunofluorescence staining for MFG-E8 and butyrophilin showed diversity in the MFG-E8-staining intensity among different fat globules in milk. HC11 cells secreted monomeric MFG-E8 with phosphatidylserine-binding activity, despite no fat globules being detected in the cells. This secretion was upregulated by not only prolactin but also by insulin or EGF. These results suggest that milk MFG-E8 was not equally present among fat globules and not necessarily intrinsic to the fat globules.

DOI： 10.1271/bbb.120412

記述言語：英語   掲載種別：研究論文（学術雑誌）  

... These results suggest that degraded but Dectin - 1 - reactive β- glucan is discharged from macrophage cells ... ROS -production inhibitors reduced soluble β- glucan discharge from the ... Soluble β- glucan produced by in vitro ROS - driven degradation activated macrophages . ...

DOI： 10.1016/j.bbrc.2012.04.009

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Microtubules form flexible fibers, which are utilized in cell proliferation and differentiation. Although the flexibility of microtubules was shown to be regulated by various microtubule-associated proteins, this regulation is still far from complete understanding. Here, we report a new potential regulator of microtubules in mammals. Gcap14 colocalizes with microtubules in mammalian cells transfected with Gcap14 expression vector. Association of Gcap14 with microtubules was confirmed by biochemical subcellular fractionation. Recombinant Gcap14 protein cosedimented with pure microtubules, indicating a direct binding between the two. Furthermore, recombinant Gcap14 was shown to have the ability of inducing microtubule bundling in vitro.

DOI： 10.1016/j.febslet.2012.04.008

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）  

DOI： 10.1242/jcs.087700

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.devcel.2011.02.001

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）  

DOI： 10.1242/dev.052605

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）  

DOI： 10.1016/j.cub.2006.06.032

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1042/BJ20051459

記述言語：英語   掲載種別：研究論文（学術雑誌）  

MFG-E8 (milk fat globule-EGF factor 8) is a peripheral membrane glycoprotein, which is expressed abundantly in lactating mammary glands and is secreted in association with fat globules. This protein consists of two-repeated EGF-like domains, a mucin-like domain and two-repeated discoidin-like domains (C-domains), and contains an integrin-binding motif (RGD sequence) in the EGF-like domain. To clarify the role of each domain on the peripheral association with the cell membrane, several domain-deletion mutants of MFG-E8 were expressed in COS-7 cells. The immunofluorescent staining of intracellular and cell-surface proteins and biochemical analyses of cell-surface-biotinylated and secreted proteins demonstrated that both of the two C-domains were required for the membrane association. During the course of these studies for domain functions, MFG-E8, but not C-domain deletion mutants, was shown to be secreted as membrane vesicle complexes. By size-exclusion chromatography and ultracentrifugation analyses, the complexes were characterized to have a high-molecular mass, low density and higher sedimentation velocity and to be detergent-sensitive. Not only such a exogenously expressed MFG-E8 but also that endogenously expressed in a mammary epithelial cell line, COMMA-1D, was secreted as the membrane vesicle-like complex. Scanning electron microscopic analyses revealed that MFG-E8 was secreted into the culture medium in association with small membrane vesicles with a size from 100 to 200 nm in diameter. Furthermore, the expression of MFG-E8 increased the number of these membrane vesicle secreted into the culture medium. These results suggest a possible role of MFG-E8 in the membrane vesicle secretion, such as budding or shedding of plasma membrane (microvesicles) and exocytosis of endocytic multivesicular bodies (exosomes).

記述言語：英語   掲載種別：研究論文（学術雑誌）  

... Biochem., 64 (4), 785-792, 2000 M An ELISA - Based Assay for Detergent - solubilized Cellular {\ss

DOI： 10.1271/bbb.64.785

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Expression of mRNA encoding MFG-E8, a milk fat-associated glycoprotein was investigated in mouse mammary gland. Two forms of mRNA, long and short variants, were shown to be expressed in the mammary tissue by RT-PCR analysis. Sequence analyses of these two variants and an isolated MFG-E8 gene segment indicated that the long and short mRNA variants resulted from an alternative splicing of a single pre-mRNA through in-flame inclusion and skipping of one exon, which encodes a proline/threonine (Pro/Thr)-rich domain. The long variant was expressed predominantly in mammary gland and the expression level was remarkably increased at late gestation and kept high during lactation. On the contrary, the short variant was detected ubiquitously in various tissues and its expression in the mammary gland was rather decreased in a lactation dependent manner. Expression of the long variant was also detected in a mouse mammary epithelial cell line, COMMA-1D, and enhanced by incubation with lactogenic hormones. Glycosylation inhibition analyses using tunicamycin and alpha-benzyl-GalNAc were conducted with COS7 cells transfected with plasmids expressing each mRNA variant, demonstrating that a fully glycosylated product of the long mRNA variant was not only N-glycosylated but also multiply O-glycosylated, whereas a product of the short one had only N-glycan(s). These results suggest that the alternative splicing plays a critical role for the mammary-specific and lactation-dependent expression of the MFG-E8 isoform and that the multiply O-glycosylated Pro/Thr-rich domain of this isoform is functionally important for formation of milk fat globules in mammary epithelial cells.

DOI： 10.1006/bbrc.1998.0107