Mucin 21/Epiglycanin Modulates Cell Adhesion
Mucin 21/Epiglycanin Modulates Cell Adhesion
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DOI：
10.1074/jbc.m109.082875
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发表时间：
2010-07-09
影响因子：
4.8
通讯作者：
Irimura, Tatsuro
中科院分区：
文献类型：
--
作者：
Yi, Yuri;Kamata-Sakurai, Mika;Irimura, Tatsuro
小鼠粘蛋白21（Muc 21）/epiglycanin的分子结构被认为具有98个15个氨基酸的串联重复序列和三个例外重复序列，每个重复序列具有12或13个氨基酸，随后是茎结构域，跨膜结构域和胞质尾。构建具有84个15个氨基酸的串联重复序列的Muc 21的cDNA，并使用Venus载体转染到HEK 293 T细胞中。被认为表达Muc 21的荧光细胞是非粘附的。当使用具有较少数目的串联重复序列的构建体时，这种抗粘附作用减弱，这表明串联重复序列结构域起着至关重要的作用。与对照细胞相比，表达Muc 21的细胞彼此粘附性和与细胞外基质组分的粘附性显著降低。抗体结合到细胞表面整合素亚基α 5，α 6和β 1的减少，在Muc 21转染在串联重复依赖性的方式，而等量的蛋白质检测通过Western印迹分析。Muc 21表达为一个大的糖蛋白，其在细胞表面被O-聚糖高度糖基化，如通过流式细胞术、蛋白质印迹和凝集素印迹检测到的。尽管至少一部分Muc 21被唾液酸化聚糖糖基化，但唾液酸的去除不影响粘附的预防。
The molecular structure of mouse Mucin 21 (Muc21)/epiglycanin is proposed to have 98 tandem repeats of 15 amino acids and three exceptional repeats with 12 or 13 amino acids each, followed by a stem domain, a transmembrane domain, and a cytoplasmic tail. A cDNA of Muc21 having 84 tandem repeats of 15 amino acids was constructed and transfected using a Venus vector into HEK 293T cells. The fluorescent cells, which were considered to express Muc21, were nonadherent. This anti-adhesion effect was lessened when constructs with smaller numbers of tandem repeats were used, suggesting that the tandem repeat domain plays a crucial role. Cells expressing Muc21 were significantly less adherent to each other and to extracellular matrix components than control cells. Antibody binding to the cell surface integrin subunits alpha 5, alpha 6, and beta 1 was reduced in Muc21 transfectants in a tandem repeat-dependent manner, whereas equal amounts of proteins were detected by Western blot analysis. Muc21 was expressed as a large glycoprotein that was highly glycosylated with O-glycans at the cell surface, as detected by flow cytometry, Western blotting, and lectin blotting. Although at least a portion of Muc21 was glycosylated with sialylated glycans, removal of sialic acid did not influence the prevention of adhesion.