Tetraspanin protein CD9 interacts with metalloprotease CD10 and enhances its release via exosomes
Tetraspanin protein CD9 interacts with metalloprotease CD10 and enhances its release via exosomes
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DOI：
10.1111/febs.12110
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发表时间：
2013-03-01
期刊：
影响因子：
5.4
通讯作者：
Filatov, Alexander
中科院分区：
文献类型：
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作者：
Mazurov, Dmitriy;Barbashova, Lubov;Filatov, Alexander
四跨膜蛋白与各种各样的跨膜和细胞内蛋白质相互作用，称为分子伴侣，并调节它们的功能。在这篇文章中，我们描述了一个新的合作伙伴的四跨膜蛋白网络，膜金属蛋白酶CD 10，这是选择性地与CD 9。通过构建四跨膜蛋白CD 9和CD 82之间的嵌合体（后者不与CD 10相互作用）或通过使用定点诱变，我们确定了从CCG基序到跨膜结构域4的大细胞外环的一部分，以及CD 9的C-末端尾部，参与与CD 10的相互作用。野生型CD 9在K562 CD 10阳性细胞中的稳定表达使外泌体释放的CD 10水平提高了5倍。相反，嵌合CD 9的表达，其中包含来自CD 82的细胞质C-末端结构域，对CD 10释放几乎没有影响。Nalm-6前B细胞中CD 9表达的短发夹RNA敲低导致微泡释放的内源性CD 10的量减少两倍。在细胞或外来体上测量的CD 10的肽酶活性与CD 10表达水平相关，并且不受CD 9表达的显著调节。我们的数据表明，CD 10与四跨膜蛋白CD 9的相互作用可以发挥重要的作用，在肽酶活性从细胞表面到外部微环境的再分配。在骨髓中，CD 10可能有助于前B细胞的成熟和B细胞向血液循环的迁移，CD 10肽酶活性与外泌体的释放可以有效地调节细胞外基质微环境。结构化数字摘要CD 9与CD 179 b、四跨膜蛋白-9、CD 63、CD 81、HLA II类、未表征蛋白C14 orf 166 B、四跨膜蛋白-14、HLA I类、半乳糖激酶、蛋白质二硫键异构酶A6、CD 98、CD 19、CD 316、CD 146、CD 92、膜联蛋白A6、CD 156 c、CD 71、CD 10、CD 29、CD 315、DAAM 1和CD 49 f抗诱饵免疫共沉淀（查看相互作用）CD 10通过抗诱饵免疫共沉淀与CD 9发生物理相互作用（查看交互：1，2，3，4）CD 10与CD 179 b、蛋白FAM 207 A、CD 9、CD 81、未表征蛋白C22 orf 13、HLA II类、未表征蛋白C3 orf 26、HLA I类物理相互作用，蛋白质C14 orf 21，Annexin A6和DAAM 1通过抗诱饵共免疫沉淀（查看相互作用）
Tetraspanins interact with a wide variety of transmembrane and intracellular proteins called molecular partners, and modulate their function. In this article, we describe a new partner of tetraspanin web, membrane metalloprotease CD10, which is selectively associated with CD9. By constructing chimeras between tetraspanins CD9 and CD82 (the latter does not interact with CD10) or by using site-directed mutagenesis, we determined that a portion of the large extracellular loop from the CCG motif to transmembrane domain4, as well as the C-terminal tail of CD9, are involved in the interaction with CD10. The stable expression of wild-type CD9 in K562 CD10-positive cells enhanced the level of CD10 released with exosomes five-fold. In contrast, the expression of chimeric CD9, which contained the cytoplasmic C-terminal domain from CD82, had little effect on CD10 release. Short hairpin RNA knockdown of CD9 expression in Nalm-6 pre-Bcells resulted in a two-fold reduction in the amount of endogenous CD10 released with microvesicles. The peptidase activity of CD10 measured either on cells or on exosomes correlated with the level of CD10 expression, and was not significantly modulated by CD9 expression as such. Our data suggest that the interaction of CD10 with tetraspanin CD9 can play an important role in the redistribution of peptidase activity from the cell surface to outer microenvironments. In bone marrow, where CD10 presumably contributes to the maturation of pre-Bcells and migration of Bcells to the blood circulation, release of CD10 peptidase activity with exosomes may effectively regulate extracellular matrix microenvironments. Structured digital abstract CD9 physically interacts with CD179b, Tetraspanin-9, CD63, CD81, HLA class II, Uncharacterized protein C14orf166B, Tetraspanin-14, HLA class I, Galactokinase, Protein disulfide-isomerase A6, CD98, CD19, CD316, CD146, CD92, Annexin A6, CD156c, CD71, CD10, CD29, CD315, DAAM1 and CD49f by anti bait coimmunoprecipitation (View interaction) CD10 physically interacts with CD9 by anti bait coimmunoprecipitation (View Interaction: 1, 2, 3, 4) CD10 physically interacts with CD179b, Protein FAM207A, CD9, CD81, Uncharacterized protein C22orf13, HLA class II, Uncharacterized protein C3orf26, HLA class I, Protein C14orf21, Annexin A6 and DAAM1 by anti bait coimmunoprecipitation (View interaction)