Proteomics

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Identification of GltB-interacting proteins by CoIP-MS

ABSTRACT: To investigate whether GltB possess more function, Co-immunoprecipitation coupled with Mass Spectrometry (CoIP-MS) was used to isolate the putative GltB binding proteins.We used strains PAO1/p-gtrS-YFP and ΔgltB/p-gtrS-YFP to ensure the membrane protein GtrS of great interest is at high levels. A target GltB-specific antibody anti-GltB was used and ΔgltB/p-gtrS-YFP was set up as a negative control. The whole gel lane was divided into small fractions and in-gel digested for liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Control sample was also analyzed in parallel to distinguish the background proteins.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Pseudomonas Aeruginosa Pao1

TISSUE(S): Cell Culture

SUBMITTER: Chenchen Xu  

LAB HEAD: Lefu Lan

PROVIDER: PXD020213 | Pride | 2021-09-09

REPOSITORIES: Pride

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Publications

Glucose-Binding of Periplasmic Protein GltB Activates GtrS-GltR Two-Component System in <i>Pseudomonas aeruginosa</i>.

Xu Chenchen C   Cao Qiao Q   Lan Lefu L  

Microorganisms 20210221 2

A two-component system GtrS-GltR is required for glucose transport activity in <i>P. aeruginosa</i> and plays a key role during <i>P. aeruginosa</i>-host interactions. However, the mechanism of action of GtrS-GltR has not been definitively established. Here, we show that <i>gltB</i>, which encodes a periplasmic glucose binding protein, is essential for the glucose-induced activation of GtrS-GltR in <i>P. aeruginosa</i>. We determined that GltB is capable of binding to membrane regulatory protein  ...[more]

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