Improved pattern for genome-based screening identifies novel cell wall-attached proteins in gram-positive bacteria
Improved pattern for genome-based screening identifies novel cell wall-attached proteins in gram-positive bacteria
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DOI：
10.1128/iai.69.6.4019-4026.2001
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发表时间：
2001-06-01
影响因子：
3.1
通讯作者：
Rasmussen, M
中科院分区：
文献类型：
--
作者：
Janulczyk, R;Rasmussen, M
随着大量测序的微生物基因组可用，用于鉴定蛋白质组或类别的工具变得越来越重要。在这里，我们提出了一种改进的模式，用于识别细胞壁附着蛋白（CWP），一组蛋白质在革兰氏阳性菌中具有不同的和重要的功能。这种三重模式是基于对65种先前描述的细胞壁附着蛋白的分析，并考虑到细胞壁分选的三个主要要求：分选酶靶区（LPXTGX）、跨膜区和带电的停止转移尾。在五种不同的革兰氏阳性菌基因组中，三重模式共鉴定出35种推定的CWP，其中19种是新的。三重模式的特异性和敏感性高于仅基于分选酶靶区域的经典模式。鉴定了几种具有非典型分选酶靶区域的推定CWP。在重要的人类病原体化脓性链球菌的完整基因组中，三方模式确定了14个推定的CWP。七个假定的S。化脓菌的蛋白质是新的，其中两个是5'核苷酸酶和支链淀粉酶。这项研究代表了第一个全基因组筛选CWP，我们得出结论，三重模式是非常适合于这一目的。使用这种模式识别CWP为研究革兰氏阳性细菌的发病机制和生理学提供了重要的可能性。
With a large number of sequenced microbial genomes available, tools for identifying groups or classes of proteins have become increasingly important. Here we present an improved pattern for the identification of cell wall-attached proteins (CWPs), a group of proteins with diverse and important functions in gram-positive bacteria. This tripartite pattern is based on analysis of 65 previously described cell wall-attached proteins and takes into account the three principal requirements for cell wall sorting; a sortase target region (LPXTGX), a membrane-spanning region, and a charged stop-transfer tail. In five different genomes of gram-positive bacteria, the tripartite pattern identified a total of 35 putative CWPs, 19 of which were novel. The specificity and sensitivity of the tripartite pattern are higher than those of the classical pattern, which is based solely on the sortase target region. Several putative CWPs with atypical sortase target regions were identified. In the complete genome of the important human pathogen Streptococcus pyogenes, the tripartite pattern identified 14 putative CWPs. Seven of the putative S. pyogenes proteins were novel, and two of these were a 5' nucleotidase and a pullulanase, This study represents the first whole-genome screening for CWPs, and we conclude that the tripartite pattern is highly suitable for this purpose. Identification of CWPs using this pattern offers important possibilities in the study of the pathogenesis and physiology of gram-positive bacteria.