Rational engineering of enzyme allosteric regulation through sequence evolution analysis.

Control of enzyme allosteric regulation is required to drive metabolic flux toward desired levels. Although the three-dimensional (3D) structures of many enzyme-ligand complexes are available, it is still difficult to rationally engineer an allosterically regulatable enzyme without decreasing its ca...

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Autores principales:	Jae-Seong Yang, Sang Woo Seo, Sungho Jang, Gyoo Yeol Jung, Sanguk Kim
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/a581ae6e26e84e8abd03716717442faa
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spelling	oai:doaj.org-article:a581ae6e26e84e8abd03716717442faa2021-11-18T05:51:10ZRational engineering of enzyme allosteric regulation through sequence evolution analysis.1553-734X1553-735810.1371/journal.pcbi.1002612https://doaj.org/article/a581ae6e26e84e8abd03716717442faa2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22807670/?tool=EBIhttps://doaj.org/toc/1553-734Xhttps://doaj.org/toc/1553-7358Control of enzyme allosteric regulation is required to drive metabolic flux toward desired levels. Although the three-dimensional (3D) structures of many enzyme-ligand complexes are available, it is still difficult to rationally engineer an allosterically regulatable enzyme without decreasing its catalytic activity. Here, we describe an effective strategy to deregulate the allosteric inhibition of enzymes based on the molecular evolution and physicochemical characteristics of allosteric ligand-binding sites. We found that allosteric sites are evolutionarily variable and comprised of more hydrophobic residues than catalytic sites. We applied our findings to design mutations in selected target residues that deregulate the allosteric activity of fructose-1,6-bisphosphatase (FBPase). Specifically, charged amino acids at less conserved positions were substituted with hydrophobic or neutral amino acids with similar sizes. The engineered proteins successfully diminished the allosteric inhibition of E. coli FBPase without affecting its catalytic efficiency. We expect that our method will aid the rational design of enzyme allosteric regulation strategies and facilitate the control of metabolic flux.Jae-Seong YangSang Woo SeoSungho JangGyoo Yeol JungSanguk KimPublic Library of Science (PLoS)articleBiology (General)QH301-705.5ENPLoS Computational Biology, Vol 8, Iss 7, p e1002612 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biology (General) QH301-705.5
spellingShingle	Biology (General) QH301-705.5 Jae-Seong Yang Sang Woo Seo Sungho Jang Gyoo Yeol Jung Sanguk Kim Rational engineering of enzyme allosteric regulation through sequence evolution analysis.
description	Control of enzyme allosteric regulation is required to drive metabolic flux toward desired levels. Although the three-dimensional (3D) structures of many enzyme-ligand complexes are available, it is still difficult to rationally engineer an allosterically regulatable enzyme without decreasing its catalytic activity. Here, we describe an effective strategy to deregulate the allosteric inhibition of enzymes based on the molecular evolution and physicochemical characteristics of allosteric ligand-binding sites. We found that allosteric sites are evolutionarily variable and comprised of more hydrophobic residues than catalytic sites. We applied our findings to design mutations in selected target residues that deregulate the allosteric activity of fructose-1,6-bisphosphatase (FBPase). Specifically, charged amino acids at less conserved positions were substituted with hydrophobic or neutral amino acids with similar sizes. The engineered proteins successfully diminished the allosteric inhibition of E. coli FBPase without affecting its catalytic efficiency. We expect that our method will aid the rational design of enzyme allosteric regulation strategies and facilitate the control of metabolic flux.
format	article
author	Jae-Seong Yang Sang Woo Seo Sungho Jang Gyoo Yeol Jung Sanguk Kim
author_facet	Jae-Seong Yang Sang Woo Seo Sungho Jang Gyoo Yeol Jung Sanguk Kim
author_sort	Jae-Seong Yang
title	Rational engineering of enzyme allosteric regulation through sequence evolution analysis.
title_short	Rational engineering of enzyme allosteric regulation through sequence evolution analysis.
title_full	Rational engineering of enzyme allosteric regulation through sequence evolution analysis.
title_fullStr	Rational engineering of enzyme allosteric regulation through sequence evolution analysis.
title_full_unstemmed	Rational engineering of enzyme allosteric regulation through sequence evolution analysis.
title_sort	rational engineering of enzyme allosteric regulation through sequence evolution analysis.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/a581ae6e26e84e8abd03716717442faa
work_keys_str_mv	AT jaeseongyang rationalengineeringofenzymeallostericregulationthroughsequenceevolutionanalysis AT sangwooseo rationalengineeringofenzymeallostericregulationthroughsequenceevolutionanalysis AT sunghojang rationalengineeringofenzymeallostericregulationthroughsequenceevolutionanalysis AT gyooyeoljung rationalengineeringofenzymeallostericregulationthroughsequenceevolutionanalysis AT sangukkim rationalengineeringofenzymeallostericregulationthroughsequenceevolutionanalysis
_version_	1718424747067310080

Rational engineering of enzyme allosteric regulation through sequence evolution analysis.

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