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Membrane and membrane‐associated proteins in Triton X‐114 extracts of Mycobacterium bovis BCG identified using a combination of gel‐based and gel‐free fractionation strategies
Author(s) -
Målen Hiwa,
Berven Frode S.,
Søfteland Tina,
Arntzen Magnus Øverlie,
D'Santos Clive S.,
De Souza Gustavo Antonio,
Wiker Harald G.
Publication year - 2008
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200700528
Subject(s) - membrane protein , integral membrane protein , membrane , mycobacterium bovis , biology , bacteria , biochemistry , peripheral membrane protein , transmembrane protein , mycobacterium tuberculosis , enzyme , chemistry , tuberculosis , medicine , genetics , receptor , pathology
Tuberculosis is an ancient disease that remains a significant global health problem. Because many membrane and membrane‐associated proteins of this pathogen represent potential targets for drugs, diagnostic probes or vaccine components, we have analysed Mycobacterium bovis , bacillus Calmette–Guérin (BCG) substrain Moreau, using Triton X‐114 for extraction of lipophilic proteins, followed by identification with LC coupled MS/MS. We identified 351 different proteins in total, and 103 (29%) were predicted as integral membrane proteins with at least one predicted transmembrane region and another 84 (23.9%) proteins had a positive grand average of hydropathicity (GRAVY) value, indicating increased probability for membrane association. Altogether 43 predicted lipoproteins (Lpps) were identified which is close to 50% of the total number of Lpps in the genome. Fifty‐four proteins, including twenty‐four predicted integral membrane proteins and seven predicted Lpps are described for the first time. The proportion of hydrophobic membrane and membrane‐associated proteins shows that Triton X‐114 is a highly efficient method for extraction of membrane proteins from bacteria, without the need for preisolation of membranes. ATP synthase, NAD(P) transhydrogenase, ubiquinone oxidoreductase and ubiquinol–cytochrome C reductase appear to represent major enzyme complexes in the membrane of Mycobacterium tuberculosis complex organisms.