Crystal structure of the outer membrane protease OmpT from Escherichia coli suggests a novel catalytic site

被引:197
作者
Vandeputte-Rutten, L
Kramer, RA
Kroon, J
Dekker, N
Egmond, MR
Gros, P
机构
[1] Univ Utrecht, Bijvoet Ctr Biomol Res, Dept Crystal & Struct Chem, NL-3584 CH Utrecht, Netherlands
[2] Univ Utrecht, Dept Enzymol & Prot Engn, Ctr Biomembranes & Lipid Enzymol, Biomembrane Inst, NL-3584 CH Utrecht, Netherlands
关键词
His-Asp dyad; lipopolysaccharide; OmpT; omptin; protease;
D O I
10.1093/emboj/20.18.5033
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
OmpT from Escherichia coli belongs to a family of highly homologous outer membrane proteases, known as omptins, which are implicated in the virulence of several pathogenic Gram-negative bacteria. Here we present the crystal structure of OmpT, which shows a 10-stranded antiparallel beta -barrel that protrudes far from the lipid bilayer into the extracellular space. We identified a putative binding site for lipopolysaccharide, a molecule that is essential for OmpT activity. The proteolytic site is located in a groove at the extracellular top of the vase-shaped beta -barrel. Based on the constellation of active site residues, we propose a novel proteolytic mechanism, involving a His-Asp dyad and an Asp-Asp couple that activate a putative nucleophilic water molecule. The active site is fully conserved within the omptin family. Therefore, the structure described here provides a sound basis for the design of drugs against omptin-mediated bacterial pathogenesis. Coordinates are in the Protein Data Bank (accession No. 1178).
引用
收藏
页码:5033 / 5039
页数:7
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