Complete alanine scanning of the Escherichia coli RbsB ribose binding protein reveals residues important for chemoreceptor signaling and periplasmic abundance.

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Serval ID
serval:BIB_5AD53A0AA296
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Complete alanine scanning of the Escherichia coli RbsB ribose binding protein reveals residues important for chemoreceptor signaling and periplasmic abundance.
Journal
Scientific reports
Author(s)
Reimer A., Maffenbeier V., Dubey M., Sentchilo V., Tavares D., Gil M.H., Beggah S., van der Meer J.R.
ISSN
2045-2322 (Electronic)
ISSN-L
2045-2322
Publication state
Published
Issued date
15/08/2017
Peer-reviewed
Oui
Volume
7
Number
1
Pages
8245
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Abstract
The Escherichia coli RbsB ribose binding protein has been used as a scaffold for predicting new ligand binding functions through in silico modeling, yet with limited success and reproducibility. In order to possibly improve the success of predictive modeling on RbsB, we study here the influence of individual residues on RbsB-mediated signaling in a near complete library of alanine-substituted RbsB mutants. Among a total of 232 tested mutants, we found 10 which no longer activated GFPmut2 reporter expression in E. coli from a ribose-RbsB hybrid receptor signaling chain, and 13 with significantly lower GFPmut2 induction than wild-type. Quantitative mass spectrometry abundance measurements of 25 mutants and wild-type RbsB in periplasmic space showed four categories of effects. Some (such as D89A) seem correctly produced and translocated but fail to be induced with ribose. Others (such as N190A) show lower induction probably as a result of less efficient production, folding and translocation. The third (such as N41A or K29A) have defects in both induction and abundance. The fourth category consists of semi-constitutive mutants with increased periplasmic abundance but maintenance of ribose induction. Our data show how RbsB modeling should include ligand-binding as well as folding, translocation and receptor binding.
Keywords
Alanine/chemistry, Alanine/metabolism, Amino Acid Substitution, Escherichia coli/genetics, Escherichia coli/metabolism, Escherichia coli Proteins/chemistry, Escherichia coli Proteins/genetics, Escherichia coli Proteins/metabolism, Models, Biological, Models, Molecular, Molecular Conformation, Molecular Imaging, Mutation, Periplasmic Binding Proteins/chemistry, Periplasmic Binding Proteins/genetics, Periplasmic Binding Proteins/metabolism, Protein Binding, Protein Transport, Ribose/metabolism, Signal Transduction
Pubmed
Web of science
Open Access
Yes
Create date
06/09/2017 11:14
Last modification date
20/08/2019 15:13
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