4.5 Article

Improved accuracy of cell surface shaving proteomics in Staphylococcus aureus using a false-positive control

期刊

PROTEOMICS
卷 10, 期 10, 页码 2037-2049

出版社

WILEY
DOI: 10.1002/pmic.200900564

关键词

Cell shaving; Microbiology; Staphylococcus aureus; Surface-exposed peptides; Surfaceome; Vaccine strategy

资金

  1. University of Southern Denmark
  2. Australian Postgraduate Award (APA)

向作者/读者索取更多资源

Proteolytic treatment of intact bacterial cells is an ideal means for identifying surface-exposed peptide epitopes and has potential for the discovery of novel vaccine targets. Cell stability during such treatment, however, may become compromised and result in the release of intracellular proteins that complicate the final analysis. Staphylococcus aureus is a major human pathogen, causing community and hospital-acquired infections, and is a serious healthcare concern due to the increasing prevalence of multiple antibiotic resistances amongst clinical isolates. We employed a cell surface shaving technique with either trypsin or proteinase-K combined with LC-MS/MS. Trypsin-derived data were controlled using a false-positive strategy where cells were incubated without protease, removed by centrifugation and the resulting supernatants digested. Peptides identified in this fraction most likely result from cell lysis and were removed from the trypsin-shaved data set. We identified 42 predicted S. aureus COL surface proteins from 260 surface-exposed peptides. Trypsin and proteinase-K digests were highly complementary with ten proteins identified by both, 16 specific to proteinase-K treatment, 13 specific to trypsin and three identified in the control. The use of a subtracted false-positive strategy improved enrichment of surface-exposed peptides in the trypsin data set to approximately 80% (124/155 peptides). Predominant surface proteins were those associated with methicillin resistance surface protein SACOL0050 (pls) and penicillin-binding protein 2' (mecA), as well as bifunctional autolysin and the extracellular matrix-binding protein Ebh. The cell shaving strategy is a rapid method for identifying surface-exposed peptide epitopes that may be useful in the design of novel vaccines against S. aureus.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.5
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据