Identification of Helicobacter pylori LolD

ABSTRACT: 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.

INSTRUMENT(S): LTQ

ORGANISM(S): Helicobacter Pylori 26695 (ncbitaxon:85962)

SUBMITTER: Mark McClain  

PROVIDER: MSV000091176 | MassIVE | Tue Jan 31 04:11:00 GMT 2023

REPOSITORIES: MassIVE