Identification of serine/threonine kinase substrates in the human pathogen group B streptococcus.
Identification of serine/threonine kinase substrates in the human pathogen group B streptococcus.
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DOI：
10.1021/pr900069n
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发表时间：
2009-05
影响因子：
4.4
通讯作者：
Rajagopal L
中科院分区：
文献类型：
--
作者：
Silvestroni A;Jewell KA;Lin WJ;Connelly JE;Ivancic MM;Tao WA;Rajagopal L
所有生物体都对它们的内部和外部环境的变化做出反应，以维持它们的生存和存在。信号传导主要通过原核生物和真核生物中蛋白质的可逆磷酸化来实现。蛋白质磷酸化状态的变化改变了其功能，从而能够控制细胞反应。许多丝氨酸/苏氨酸激酶调节真核生物的细胞反应。虽然在真核生物中很常见，但丝氨酸/苏氨酸激酶直到最近才在原核生物中被鉴定。我们已经描述了人类病原体B组链球菌（GBS，无乳链球菌）编码一个单一的膜相关的丝氨酸/苏氨酸激酶（Stk 1），这对该细菌的毒力很重要。本研究采用磷酸肽富集与质谱联用技术对GBS中丝氨酸（S）和苏氨酸（T）磷酸肽进行富集和鉴定。从Stk 1表达菌株鉴定的S/T磷酸肽与同基因stk 1突变体的比较表明，10种蛋白质是GBS Stk 1酶的潜在底物。这些蛋白质中的一些在体外被Stk 1磷酸化，并且将磷酸化的苏氨酸定点取代为丙氨酸消除了Stk 1底物的磷酸化。总的来说，这些研究提供了一种新的方法来识别丝氨酸/苏氨酸激酶底物，以深入了解它们在人类病原体如GBS中的信号传导。
All living organisms respond to changes in their internal and external environment for their survival and existence. Signaling is primarily achieved through reversible phosphorylation of proteins in both prokaryotes and eukaryotes. A change in the phosphorylation state of a protein alters its function to enable the control of cellular responses. A number of serine/threonine kinases regulate the cellular responses of eukaryotes. Although common in eukaryotes, serine/threonine kinases have only recently been identified in prokaryotes. We have described that the human pathogen Group B Streptococcus (GBS, Streptococcus agalactiae) encodes a single membrane-associated, serine/threonine kinase (Stk1) that is important for virulence of this bacterium. In this study, we used a combination of phosphopeptide enrichment and mass spectrometry to enrich and identify serine (S) and threonine (T) phosphopeptides of GBS. A comparison of S/T phosphopeptides identified from the Stk1 expressing strains to the isogenic stk1 mutant indicates that 10 proteins are potential substrates of the GBS Stk1 enzyme. Some of these proteins are phosphorylated by Stk1 in vitro and a site-directed substitution of the phosphorylated threonine to an alanine abolished phosphorylation of an Stk1 substrate. Collectively, these studies provide a novel approach to identify serine/threonine kinase substrates for insight into their signaling in human pathogens like GBS.
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影响因子：
3.6
作者：
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