Elucidation of the substrate specificity of the MASP-2 protease of the lectin complement pathway and identification of the enzyme as a major physiological target of the serpin, C1-inhibitor

被引:59
作者
Kerr, Felicity K.
Thomas, Adele R.
Wijeyewickrema, Lakshmi C.
Whisstock, James C.
Boyd, Sarah E.
Kaiserman, Dion
Matthews, Antony Y.
Bird, Phillip I.
Thielens, Nicole M.
Rossi, Veronique
Pike, Robert N. [1 ]
机构
[1] Monash Univ, Dept Biochem & Mol Biol, Clayton, Vic 3800, Australia
[2] Monash Univ, ARC Ctr Struct & Funct Microbiol Genom, Clayton, Vic 3800, Australia
[3] Monash Univ, Fac Technol & Informat, Clayton, Vic 3800, Australia
[4] Inst Biol Struct Jean Pierre Ebel, Lab Enzymol Mol, Grenoble, France
基金
英国医学研究理事会;
关键词
mannose-binding lectin-associated serine protease-2; complement; active site specificity; C1s; C1-inhibitor; serpin; protease;
D O I
10.1016/j.molimm.2007.07.008
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
Complement is a central component of host defence, but unregulated activation can contribute to disease. The system can be initiated by three pathways: classical, alternative and lectin. The classical and lectin pathways are initiated by the C1 and mannose-binding lectin (MBL) or ficolin complexes, respectively, with C1s the executioner protease of the C1 complex and MASP-2 its counterpart in the lectin complexes. These proteases in turn cleave the C4 and C2 components of the system. Here we have elucidated the cleavage specificity of MASP-2 using a randomised substrate phage display library. Apart from the crucial P, position, the MASP-2 S-2 and S-3 subsites (in that order) play the greatest role in determining specificity, with Gly residues preferred at P-2 and Leu or hydrophobic residues at P-3-Cleavage of peptide substrates representing the known physiological cleavage sequences in C2, C4 or the serpin C1-inhibitor (a likely regulator of MASP-2) revealed that MASP-2 is up to 1000 times more catalytically active than C1s. C1-inhibitor inhibited MASP-2 50-fold faster than C1s and much faster than any other protease tested to date, implying that MASP-2 is a major physiological target of C1-inhibitor. (c) 2007 Elsevier Ltd. All rights reserved.
引用
收藏
页码:670 / 677
页数:8
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