Proteolytic cleavage of urokinase-type plasminogen activator by stromelysin-1 (MMP-3)

被引:60
作者
Ugwu, F
Van Hoef, B
Bini, A
Collen, D
Lijnen, HR
机构
[1] Catholic Univ Louvain, Ctr Mol & Vasc Biol, B-3000 Louvain, Belgium
[2] New York Blood Ctr, Lindsley F Kimball Res Inst, Lab Blood Coagulat Biochem, New York, NY 10021 USA
关键词
D O I
10.1021/bi9728708
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Matrix metalloproteinase-3 (MMP-3, or stromelysin-1) specifically hydrolyzes the Glu(143)-Leu(144) peptide bond in 45-kDa single-chain urokinase-type plasminogen activator (scu-PA) and in its two-chain (tcu-PA) derivative, yielding a 17-kDa NH2-terminal domain comprising the u-PA receptor (u-PAR) binding site and a 32-kDa COOH-terminal moiety containing the serine proteinase domain of u-PA. The conversion is completely abolished in the presence of the MMP inhibitors EDTA or 1,10-phenanthroline. Biospecific interaction analysis indicates that binding of MMP-3 occurs through the 32-kDa fragment. The 32-kDa fragment derived from scu-PA (scu-PA-32k) has a specific activity of less than or equal to 500 IU/mg, but it can be activated with plasmin to a two-chain derivative (tcu-PA-32k) with a specific activity of 79 000 IU/mg, tcu-PA and tcu-PA-32k moieties derived from scu-PA-32k by plasmin or from tcu-PA by MMP-3 have comparable amidolytic activities toward the chromogenic substrate S-2444 (k(cat)/K-m of 110 and 160 mM(-1) s(-1), respectively) and similar plasminogen activating activities in a coupled chromogenic substrate assay. Specific binding of the 17-kDa NH2-terminal domain to THP-1 monocytoid cells is completely abolished by competition with scu-PA but is not affected by scu-PA-32k (residual binding of 88 +/- 9% (mean +/- SEM; n = 3) with 25-fold molar excess). Thus, MMP-3 removes a functional NH2-terminal u-PAR-binding domain from u-PA without affecting its enzymatic properties.
引用
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页码:7231 / 7236
页数:6
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