ADAM13 disintegrin and cysteine-rich domains bind to the second heparin-binding domain of fibronectin
ADAM13 disintegrin and cysteine-rich domains bind to the second heparin-binding domain of fibronectin
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DOI：
10.1074/jbc.m201792200
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发表时间：
2002-06-28
影响因子：
4.8
通讯作者：
Alfandari, D
中科院分区：
文献类型：
--
作者：
Gaultier, A;Cousin, H;Alfandari, D
ADAM13是解整合素金属蛋白酶蛋白家族的成员，在颅神经嵴细胞表面表达，对其迁移至关重要。ADAM13是一种活性蛋白酶，在体外能切割纤连蛋白，在体内能重塑纤连蛋白底物。利用一种包含ADAM13的解整合素结构域和富含半胱氨酸结构域的重组分泌蛋白，我们表明该蛋白的这个“黏附”区域直接与纤连蛋白结合。对应各种功能结构域的纤连蛋白融合蛋白被用于确定第二个肝素结合结构域为ADAM13结合位点。该结构域内 syndecan结合位点（PPRR→PPTM）的突变消除了ADAM13的重组解整合素和富含半胱氨酸结构域的结合。我们进一步表明，ADAM13的黏附性解整合素和富含半胱氨酸结构域可通过β1整合素促进细胞黏附。这种黏附需要整合素激活，并且可被针对ADAM13富含半胱氨酸结构域和β1整合素的抗体所阻断。最后，野生型ADAM13（而非其金属蛋白酶结构域的E/A突变体）可从细胞表面脱落，释放与解整合素和富含半胱氨酸结构域相关联的金属蛋白酶结构域。这表明ADAM13的脱落可能涉及其自身的金属蛋白酶活性，并且释放的蛋白酶可能与整合素和细胞外基质蛋白相互作用。
ADAM13 is a member of the disintegrin and metalloprotease protein family that is expressed on cranial neural crest cells surface and is essential for their migration. ADAM13 is an active protease that can cleave fibronectin in vitro and remodel a fibronectin substrate in vivo. Using a recombinant secreted protein containing both disintegrin and cysteine-rich domains of ADAM13, we show that this "adhesive" region of the protein binds directly to fibronectin. Fibronectin fusion proteins corresponding to the various functional domains were used to define the second heparin-binding domain as the ADAM13 binding site. Mutation of the syndecan-binding site (PPRR --> PPTM) within this domain abolishes binding of the recombinant disintegrin and cysteine-rich domains of ADAM13. We further show that the adhesive disintegrin and cysteine-rich domain of ADAM13 can promote cell adhesion via P, integrins. This adhesion requires integrin activation and can be prevented by antibodies to the cysteine-rich domain of ADAM13 and P, integrin. Finally, wild type, but not the E/A mutant of ADAM13 metalloprotease domain, can be shed from the cell surface, releasing the metalloprotease domain associated with the disintegrin and cysteine-rich domains. This suggests that ADAM13 shedding may involve its own metalloprotease activity and that the released protease may interact with both integrins and extracellular matrix proteins.