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

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 influ...

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Autores principales:	Artur Reimer, Vitali Maffenbeier, Manupriyam Dubey, Vladimir Sentchilo, Diogo Tavares, Manuel Hernandez Gil, Siham Beggah, Jan Roelof van der Meer
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0b1e4619ab914a33b84f04afc5cf2d7e
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spelling	oai:doaj.org-article:0b1e4619ab914a33b84f04afc5cf2d7e2021-12-02T12:31:53ZComplete alanine scanning of the Escherichia coli RbsB ribose binding protein reveals residues important for chemoreceptor signaling and periplasmic abundance10.1038/s41598-017-08035-52045-2322https://doaj.org/article/0b1e4619ab914a33b84f04afc5cf2d7e2017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08035-5https://doaj.org/toc/2045-2322Abstract 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.Artur ReimerVitali MaffenbeierManupriyam DubeyVladimir SentchiloDiogo TavaresManuel Hernandez GilSiham BeggahJan Roelof van der MeerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Artur Reimer Vitali Maffenbeier Manupriyam Dubey Vladimir Sentchilo Diogo Tavares Manuel Hernandez Gil Siham Beggah Jan Roelof van der Meer Complete alanine scanning of the Escherichia coli RbsB ribose binding protein reveals residues important for chemoreceptor signaling and periplasmic abundance
description	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.
format	article
author	Artur Reimer Vitali Maffenbeier Manupriyam Dubey Vladimir Sentchilo Diogo Tavares Manuel Hernandez Gil Siham Beggah Jan Roelof van der Meer
author_facet	Artur Reimer Vitali Maffenbeier Manupriyam Dubey Vladimir Sentchilo Diogo Tavares Manuel Hernandez Gil Siham Beggah Jan Roelof van der Meer
author_sort	Artur Reimer
title	Complete alanine scanning of the Escherichia coli RbsB ribose binding protein reveals residues important for chemoreceptor signaling and periplasmic abundance
title_short	Complete alanine scanning of the Escherichia coli RbsB ribose binding protein reveals residues important for chemoreceptor signaling and periplasmic abundance
title_full	Complete alanine scanning of the Escherichia coli RbsB ribose binding protein reveals residues important for chemoreceptor signaling and periplasmic abundance
title_fullStr	Complete alanine scanning of the Escherichia coli RbsB ribose binding protein reveals residues important for chemoreceptor signaling and periplasmic abundance
title_full_unstemmed	Complete alanine scanning of the Escherichia coli RbsB ribose binding protein reveals residues important for chemoreceptor signaling and periplasmic abundance
title_sort	complete alanine scanning of the escherichia coli rbsb ribose binding protein reveals residues important for chemoreceptor signaling and periplasmic abundance
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/0b1e4619ab914a33b84f04afc5cf2d7e
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Complete alanine scanning of the Escherichia coli RbsB ribose binding protein reveals residues important for chemoreceptor signaling and periplasmic abundance

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