Anchoring of surface proteins to the cell wall of Staphylococcus aureus -: Sortase catalyzed in vitro transpeptidation reaction using LPXTG peptide and NH2-Gly3 substrates
Anchoring of surface proteins to the cell wall of Staphylococcus aureus -: Sortase catalyzed in vitro transpeptidation reaction using LPXTG peptide and NH2-Gly3 substrates
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DOI：
10.1074/jbc.275.13.9876
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发表时间：
2000-03-31
影响因子：
4.8
通讯作者：
Schneewind, O
中科院分区：
文献类型：
--
作者：
Ton-That, H;Mazmanian, SK;Schneewind, O
金黄色葡萄球菌分类酶通过裂解LPXTG基序上的多肽将表面蛋白锚定到细胞壁被膜上，表面蛋白通过C端羧基和五甘氨酸交叉桥的氨基之间的酰胺键连接到肽聚糖上。我们发现，在NH2-Gly(3)的存在下，苏氨酸和甘氨酸之间的肽键上含有LPXTG：基序的重组索糖酶水解肽，只催化转肽反应，将苏氨酸的羧基连接到NH2-Gly(3)的氨基上。在氨基供体的存在下，LPXTG基序的山梨酸酶介导的切割速率增加。半胱氨酸184的水解和转肽需要半胱氨酸184的巯基，这表明山梨酸酶通过形成硫酯酰基酶中间体来催化表面蛋白锚定的转肽反应。
Staphylococcus aureus sortase anchors surface proteins to the cell wall envelope by cleaving polypeptides at the LPXTG motif, Surface proteins are linked to the peptidoglycan by an amide bond between the C-terminal carboxyl and the amino group of the pentaglycine cross bridge. We find that purified recombinant sortase hydrolyzed peptides bearing an LPXTG: motif at the peptide bond between threonine and glycine, In the presence of NH2-Gly(3), sortase catalyzed exclusively a transpeptidation reaction, linking the carboxyl group of threonine to the amino group of NH2-Gly(3). In the presence of amino group donors the rate of sortase mediated cleavage at the LPXTG motif was increased. Hydrolysis and transpeptidation required the sulfhydryl of cysteine 184, suggesting that sortase catalyzed the transpeptidation reaction of surface protein anchoring via the formation of a thioester acyl-enzyme intermediate.