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运用基于数据非依赖采集(SWATH)的蛋白质组学方法理解人类胃部病原体幽门螺杆菌的双态生活方式。

Understanding the dimorphic lifestyles of human gastric pathogen Helicobacter pylori using the SWATH-based proteomics approach.

作者信息

Loke Mun Fai, Ng Chow Goon, Vilashni Yeespana, Lim Justin, Ho Bow

机构信息

Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.

Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.

出版信息

Sci Rep. 2016 May 25;6:26784. doi: 10.1038/srep26784.

Abstract

Helicobacter pylori may reside in the human stomach as two morphological forms: the culturable spiral form and the viable but non-culturable (VBNC) coccoid form. This bacterium transforms from spiral to coccoid under in vitro suboptimal conditions. However, both spiral and coccoid have demonstrated its infectivity in laboratory animals, suggesting that coccoid may potentially be involved in the transmission of H. pylori. To determine the relevance of the coccoid form in viability and infectivity, we compared the protein profiles of H. pylori coccoids obtained from prolonged (3-month-old) culture with that of 3-day-old spirals of two H. pylori standard strains using SWATH (Sequential Window Acquisition of all Theoretical mass spectra)-based approach. The protein profiles reveal that the coccoids retained basal level of metabolic proteins and also high level of proteins that participate in DNA replication, cell division and biosynthesis demonstrating that coccoids are viable. Most interestingly, these data also indicate that the H. pylori coccoids possess higher level of proteins that are involved in virulence and carcinogenesis than their spiral counterparts. Taken together, these findings have important implications in the understanding on the pathogenesis of H. pylori-induced gastroduodenal diseases, as well as the probable transmission mode of this bacterium.

摘要

幽门螺杆菌可能以两种形态存在于人的胃部

可培养的螺旋形和活的但不可培养的(VBNC)球形。在体外非最适条件下,这种细菌会从螺旋形转变为球形。然而,螺旋形和球形在实验动物中均已证明具有感染性,这表明球形可能潜在地参与了幽门螺杆菌的传播。为了确定球形形态在生存能力和感染性方面的相关性,我们使用基于SWATH(所有理论质谱的顺序窗口采集)的方法,比较了从长期(3个月大)培养获得的幽门螺杆菌球形与两种幽门螺杆菌标准菌株3天大的螺旋形的蛋白质谱。蛋白质谱显示,球形保留了基础水平的代谢蛋白,以及参与DNA复制、细胞分裂和生物合成的高水平蛋白质,这表明球形是有活力的。最有趣的是,这些数据还表明,与螺旋形相比,幽门螺杆菌球形具有更高水平的参与毒力和致癌作用的蛋白质。综上所述,这些发现对于理解幽门螺杆菌引起的胃十二指肠疾病的发病机制以及这种细菌可能的传播方式具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a95c/4879699/21459015ab24/srep26784-f1.jpg

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