Improving basic and membrane protein MS detection of the culture filtrate proteins from Mycobacterium tuberculosis H37Rv by biomimetic affinity prefractionation

Guorong Ma; Kang Zhang; Fuqiang Zhou; Lina Gao; Zhanqiang Sun; Li Deng; Shulin Zhang; Rongxiu Li

doi:10.1002/pmic.201600177

Improving basic and membrane protein MS detection of the culture filtrate proteins from Mycobacterium tuberculosis H37Rv by biomimetic affinity prefractionation

Proteomics. 2017 May;17(10):e1600177. doi: 10.1002/pmic.201600177.

Authors

Guorong Ma¹, Kang Zhang¹, Fuqiang Zhou¹, Lina Gao², Zhanqiang Sun³, Li Deng⁴, Shulin Zhang³, Rongxiu Li^{1

5}

Affiliations

¹ State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, P. R. China.
² Clinical Laboratory of the Second Affiliated Hospital, Lanzhou University, Lanzhou, P. R. China.
³ Department of Immunology and Microbiology, School of Medicine, Shanghai Jiao Tong University, Shanghai, P. R. China.
⁴ Shanghai HyCharm Inc., Shanghai, P. R. China.
⁵ Engineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, Shanghai, P. R. China.

PMID: 28393466
DOI: 10.1002/pmic.201600177

Abstract

The culture filtrate proteins (CFPs) from Mycobacterium tuberculosis have been shown to induce protective immune responses in human and animal models, making them a promising source of candidate targets for tuberculosis drugs, vaccines, and diagnostics. The constituents of the M. tuberculosis CFP proteome are complex and vary with growth conditions. To effectively profile CFPs, gel-based prefractionation is usually performed before MS analysis. In this study, we describe a novel prefractionation approach by which the proteome is divided into seven partially overlapping fractions by biomimetic affinity chromatography (BiAC) using a six-column cascade. The LC-MS/MS analysis of individual fractions identified a total of 541 CFPs, including 61 first-time identifications. Notably, ∼1/3 (20/61) of these novel CFPs are membrane proteins, among which nine proteins have 2-14 transmembrane domains. In addition, ∼1/4 (14/61) of the CFPs are basic proteins with pI values greater than 9.0. Our data demonstrate that biomimetic affinity chromatography prefractionation markedly improves protein detection by LC-MS/MS, and the coverage of basic and hydrophobic proteins in particular is remarkably increased.

Keywords: Biomimetic affinity chromatography; Culture filtrate proteins; LC-MS/MS; Mycobacterium tuberculosis.

Abstract

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