Crystal structure of PBP2x from a highly penicillin-resistant Streptococcus pneumoniae clinical isolate - A mosaic framework containing 83 mutations

被引:95
作者
Dessen, A
Mouz, N
Gordon, E
Hopkins, J
Dideberg, O
机构
[1] Inst Biol Struct Jean Pierre Ebel, Lab Cristallog Macromol, CNRS, CEA, F-38027 Grenoble, France
[2] Inst Biol Struct Jean Pierre Ebel, Lab Ingn Macromol, CNRS, CEA, F-38027 Grenoble, France
关键词
D O I
10.1074/jbc.M107608200
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
Penicillin-binding proteins (PBPs) are the main targets for beta -lactam antibiotics, such as penicillins and cephalosporins, in a wide range of bacterial species. In some Gram-positive strains, the surge of resistance to treatment with beta -lactams is primarily the result of the proliferation of mosaic PBP-encoding genes, which encode novel proteins by recombination. PBP2x is a primary resistance determinant in Streptococcus pneumoniae, and its modification is an essential step in the development of high level beta -lactam resistance. To understand such a resistance mechanism at an atomic level, we have solved the x-ray crystal structure of PBP2x from a highly penicillin-resistant clinical isolate of S. pneumoniae, Sp328, which harbors 83 mutations in the soluble region. In the proximity of the Sp328 PBP2x* active site, the Thr(338)--> Ala mutation weakens the local hydrogen bonding network, thus abrogating the stabilization of a crucial buried water molecule. In addition, the Ser(389)--> Leu and Asn(514)--> His mutations produce a destabilizing effect that generates an "open" active site. It has been suggested that peptidoglycan substrates for beta -lactam-resistant PBPs contain a large amount of abnormal, branched peptides, whereas sensitive strains tend to catalyze cross-linking of linear forms. Thus, in vivo, an "open" active site could facilitate the recognition of distinct, branched physiological substrates.
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页码:45106 / 45112
页数:7
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