Priority changes between biofilm exopolysaccharides synthesis and rhamnolipids production are mediated by a c-di-GMP-specific phosphodiesterase NbdA in Pseudomonas aeruginosa

Liu, Shixun; Xu, Anming; Xie, Bin; Xin, Fengxue; Dong, Weiliang; Zhou, Jie; Jiang, Min

doi:10.1016/j.isci.2022.105531

Cited by 3 publications

(1 citation statement)

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“…NbdA has been reported to be involved in biofilm dispersal, swimming motility, and rhamnolipid production of P. aeruginosa (Li et al 2013 , Xin et al 2019 , Liu et al 2022 ). These functions are directly related to the nucleotide second messenger c-di-GMP and the enzymatic activity of NbdA.…”

Section: Discussionmentioning

confidence: 99%

Characterization of a soluble library of thePseudomonas aeruginosaPAO1 membrane proteome with emphasis on c-di-GMP turnover enzymes

Scherhag,

Räschle,

Unbehend

et al. 2023

microLife

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Studies of protein–protein interactions in membranes are very important to fully understand the biological function of a cell. The extraction of proteins from the native membrane environment is a critical step in the preparation of membrane proteins that but might affect the stability of protein complexes. In this work, we used the amphiphilic diisobutylene/maleic acid copolymer (DIBMA) to extract the membrane proteome of the opportunistic pathogen Pseudomonas aeruginosa, thereby creating a soluble membrane-protein library within a native-like lipid-bilayer environment. Size fractionation of nanodisc-embedded proteins and subsequent mass spectrometry enabled the identification of 3358 proteins. The native membrane-protein library showed a very good overall coverage compared to previous proteome data. The pattern of size fractionation indicated that protein complexes were preserved in the library. More than 20 previously described complexes, e.g., the SecYEG and Pili complexes, were identified and analyzed for coelution. Although the mass-spectrometric dataset alone did not reveal new protein complexes, combining pulldown assays with mass spectrometry was successful in identifying new protein interactions in the native membrane-protein library. Thus, we identified several candidate proteins for interactions with the membrane phosphodiesterase NbdA, a member of the c-di-GMP network. We confirmed the candidate proteins CzcR, PA4200, SadC and PilB as novel interaction partners of NbdA using the bacterial adenylate cyclase two-hybrid assay. Taken together, this work demonstrates the usefulness of the native membrane-protein library of P. aeruginosa for the investigation of protein interactions and membrane-protein complexes. Data are available via ProteomeXchange with identifier PXD039702 and PXD039700.

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Section: Discussionmentioning

confidence: 99%