Luu Laurence Don Wai, Octavia Sophie, Zhong Ling, Raftery Mark, Sintchenko Vitali, Lan Ruiting
School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales, Australia.
Mark Wainwright Analytical Centre, University of New South Wales, Sydney, New South Wales, Australia.
J Proteomics. 2017 Mar 31;158:43-51. doi: 10.1016/j.jprot.2017.02.010. Epub 2017 Feb 24.
Our understanding of the Bordetella pertussis secretome remains limited including the role of different growth conditions in the secretome. In this study the secretome of L1423, a clinical isolate from the 2008-2012 Australian epidemic, cultured on Stainer-Scholte (SS) and Thalen-IJssel (THIJS) media for 12h was characterised using liquid chromatography-mass spectrometry (LC-MS/MS). In the supernatant, LC-MS/MS identified 260 proteins with 143 bioinformatically predicted to be secreted. Eighty percent of proteins were identified in both media. Proteins secreted were functionally associated with cell surface (41%), pathogenicity (16%) and transport (17%). The most abundant proteins identified were pathogenic proteins including toxins (PtxA and CyaA), adhesins (TcfA) and type III secretion (T3SS) proteins. There were 46 proteins found uniquely in THIJS including 8 virulence associated proteins. These included T3SS proteins, adhesins (FhaL and FhaS) and a putative toxin (BP1251). Nine proteins were found uniquely in SS and these were metabolic and transport-related proteins. None of the unique proteins detected in SS were known to be virulence associated. This study found that THIJS promotes secretion of virulence factors based on the number of unique virulence proteins found and may be a growth media of choice for the study of B. pertussis virulence and vaccine development.
Over the past two decades, the number of B. pertussis notifications has risen despite vaccination. There is a greater need to understand the biology behind B. pertussis infections. The secretome of B. pertussis in two different media was characterised using LC-MS/MS. The results showed that THIJS promotes secretion of importance virulence factors which may be important for the development of vaccines.
我们对百日咳博德特氏菌分泌组的了解仍然有限,包括不同生长条件在分泌组中的作用。在本研究中,使用液相色谱 - 质谱联用(LC-MS/MS)对一株来自2008 - 2012年澳大利亚疫情的临床分离株L1423在施泰纳 - 肖尔特(SS)和塔伦 - 艾瑟尔(THIJS)培养基上培养12小时后的分泌组进行了表征。在上清液中,LC-MS/MS鉴定出260种蛋白质,其中143种经生物信息学预测为分泌蛋白。两种培养基中均鉴定出80%的蛋白质。分泌的蛋白质在功能上与细胞表面(41%)、致病性(16%)和转运(17%)相关。鉴定出的最丰富的蛋白质是致病蛋白,包括毒素(PtxA和CyaA)、黏附素(TcfA)和III型分泌(T3SS)蛋白。在THIJS中独特发现了46种蛋白质,其中包括8种与毒力相关的蛋白质。这些包括T3SS蛋白、黏附素(FhaL和FhaS)和一种假定毒素(BP1251)。在SS中独特发现了9种蛋白质,这些是与代谢和转运相关的蛋白质。在SS中检测到的独特蛋白质均未知与毒力相关。本研究发现,基于发现的独特毒力蛋白数量,THIJS促进毒力因子的分泌,可能是研究百日咳博德特氏菌毒力和疫苗开发的首选生长培养基。
在过去二十年中,尽管进行了疫苗接种,百日咳博德特氏菌的报告病例数仍有所上升。更需要了解百日咳博德特氏菌感染背后的生物学机制。使用LC-MS/MS对百日咳博德特氏菌在两种不同培养基中的分泌组进行了表征。结果表明,THIJS促进重要毒力因子的分泌,这可能对疫苗开发很重要。