We engineered a derivative of H. pylori strain 26695 that expresses an N-terminal DDK-tagged version of LolF from the endogenous locus. As controls, we also engineered strains to express DDK-tagged versions of two unrelated inner membrane proteins ExbD2 and FrdC. The tagged proteins were affinity purified and co-purifying proteins were identified by LC/MS-MS. We then performed an unbiased analysis using SAINTexpress (Significance Analysis of INTeractome) that included data from affinity purifications of test (i.e., LolF-DDK) and control (i.e., ExbD2-DDK and FrdC-DDK) bait proteins. This led to identification of HP0179 as an interaction partner of LolF. We then engineered a derivative of H. pylori strain 26695 that expresses a C-terminal HA-tagged version of HP0179. As controls, additional strains were generated that express HA-tagged versions of two unrelated proteins HP0838 and CagF. he tagged proteins were affinity purified and co-purifying proteins were identified by LC/MS-MS. We then performed an unbiased analysis using SAINTexpress that included data from affinity purifications of test (i.e., HP0179-HA) and control (i.e., HP0838-HA and CagF-HA) bait proteins. This led to identification of LolF as an interaction partner of HP0179.
[doi:10.25345/C5BZ61J5J]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: Helicobacter pylori ; lipoprotein ; posttranslational protein modification ; Affinity purification-Mass spectrometry ; ATP-Binding Cassette Transporters ; ATP-Binding Cassette ATP-binding proteins
Principal Investigators: (in alphabetical order) |
Mark McClain, Vanderbilt University Medical Cente, United States |
Submitting User: | mmcclain |
McClain MS, Bryant KN, McDonald WH, Algood HMS, Cover TL.
Identification of an Essential LolD-Like Protein in Helicobacter pylori.
J Bacteriol. Epub 2023 Mar 27.
McClain, M.S., K.N. Bryant, W.H. McDonald, H.M.S. Algood, and T.L. Cover.
Identification of an essential LolD-like protein in Helicobacter pylori.
Journal of Bacteriology, In Press.
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