Identification of sortase substrates by specificity profiling
Identification of sortase substrates by specificity profiling
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DOI：
10.1016/j.bmc.2017.06.033
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发表时间：
2017-09-15
影响因子：
3.5
通讯作者：
Schwarzer, Dirk
中科院分区：
文献类型：
--
作者：
Schmohl, Lena;Bierlmeier, Jan;Schwarzer, Dirk
分选酶催化表面蛋白与革兰氏阳性细菌肽聚糖层的附着，进一步成为蛋白质化学的有力工具。在催化过程中，分选酶裂解含有LPxTG （x =任何氨基酸）分选基序的供体底物，形成酶结合的硫酯，并将该中间体连接到含有n端甘氨酸残基的受体蛋白上。除了确定的金黄色葡萄球菌的分类酶A外，该酶的几个同源物已在革兰氏阳性细菌的基因组中被鉴定出来。我们分析了葡萄球菌和链球菌起源的7种分类酶的特异性，并观察到后一类分类酶对供体和受体底物的特异性比其葡萄球菌对应物更宽松。与规范的分类基序LPKTG相比，链球菌分选酶更倾向于LPKLG供体底物序列。这些发现可能有助于利用链球菌分选酶作为蛋白质化学的工具。(C) 2017 Elsevier Ltd.版权所有。
Sortases catalyze the attachment of surface proteins to the peptidoglycan layer of gram-positive bacteria and further represent powerful tools of protein chemistry. During catalysis sortases cleave a donor substrate containing the LPxTG (x = any amino acid) sorting motif under formation of an enzyme-bound thioester and ligate this intermediate to an acceptor protein containing an N-terminal glycine residue. In addition to the well-established sortase A of Staphylococcus aureus several homologs of this enzyme have been identified in the genomes of gram-positive bacteria. We have profiled the specificity of seven sortases of Staphylococci and Streptococci origin and observed that sortases of the latter class displayed a more relaxed specificity for donor and acceptor substrates than their Staphylococci counterparts. Streptococci sortases prefer an LPKLG donor substrate sequence compared to the canonical sorting motif LPKTG. These findings might facilitate the use of Streptococci sortases as tools of protein chemistry. (C) 2017 Elsevier Ltd. All rights reserved.