Evolution of allosteric citrate binding sites on 6-phosphofructo-1-kinase.

As an important part of metabolism, metabolic flux through the glycolytic pathway is tightly regulated. The most complex control is exerted on 6-phosphofructo-1-kinase (PFK1) level; this control overrules the regulatory role of other allosteric enzymes. Among other effectors, citrate has been report...

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Autores principales:	Aleksandra Usenik, Matic Legiša
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/15018ccb70c943c6ad00ac5e1b28eb4b
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spelling	oai:doaj.org-article:15018ccb70c943c6ad00ac5e1b28eb4b2021-11-18T07:36:37ZEvolution of allosteric citrate binding sites on 6-phosphofructo-1-kinase.1932-620310.1371/journal.pone.0015447https://doaj.org/article/15018ccb70c943c6ad00ac5e1b28eb4b2010-11-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21124851/?tool=EBIhttps://doaj.org/toc/1932-6203As an important part of metabolism, metabolic flux through the glycolytic pathway is tightly regulated. The most complex control is exerted on 6-phosphofructo-1-kinase (PFK1) level; this control overrules the regulatory role of other allosteric enzymes. Among other effectors, citrate has been reported to play a vital role in the suppression of this enzyme's activity. In eukaryotes, amino acid residues forming the allosteric binding site for citrate are found both on the N- and the C-terminal region of the enzyme. These site has evolved from the phosphoenolpyruvate/ADP binding site of bacterial PFK1 due to the processes of duplication and tandem fusion of prokaryotic ancestor gene followed by the divergence of the catalytic and effector binding sites. Stricter inhibition of the PFK1 enzyme was needed during the evolution of multi-cellular organisms, and the most stringent control of PFK1 by citrate occurs in vertebrates. By substituting a single amino acid (K557R or K617A) as a component of the allosteric binding site in the C-terminal region of human muscle type PFK-M with a residue found in the corresponding site of a fungal enzyme, the inhibitory effect of citrate was attenuated. Moreover, the proteins carrying these single mutations enabled growth of E. coli transformants encoding mutated human PFK-M in a glucose-containing medium that did not support the growth of E. coli transformed with native human PFK-M. Substitution of another residue at the citrate-binding site (D591V) of human PFK-M resulted in the complete loss of activity. Detailed analyses revealed that the mutated PFK-M subunits formed dimers but were unable to associate into the active tetrameric holoenzyme. These results suggest that stricter control over glycolytic flux developed in metazoans, whose somatic cells are largely characterized by slow proliferation.Aleksandra UsenikMatic LegišaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 11, p e15447 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Aleksandra Usenik Matic Legiša Evolution of allosteric citrate binding sites on 6-phosphofructo-1-kinase.
description	As an important part of metabolism, metabolic flux through the glycolytic pathway is tightly regulated. The most complex control is exerted on 6-phosphofructo-1-kinase (PFK1) level; this control overrules the regulatory role of other allosteric enzymes. Among other effectors, citrate has been reported to play a vital role in the suppression of this enzyme's activity. In eukaryotes, amino acid residues forming the allosteric binding site for citrate are found both on the N- and the C-terminal region of the enzyme. These site has evolved from the phosphoenolpyruvate/ADP binding site of bacterial PFK1 due to the processes of duplication and tandem fusion of prokaryotic ancestor gene followed by the divergence of the catalytic and effector binding sites. Stricter inhibition of the PFK1 enzyme was needed during the evolution of multi-cellular organisms, and the most stringent control of PFK1 by citrate occurs in vertebrates. By substituting a single amino acid (K557R or K617A) as a component of the allosteric binding site in the C-terminal region of human muscle type PFK-M with a residue found in the corresponding site of a fungal enzyme, the inhibitory effect of citrate was attenuated. Moreover, the proteins carrying these single mutations enabled growth of E. coli transformants encoding mutated human PFK-M in a glucose-containing medium that did not support the growth of E. coli transformed with native human PFK-M. Substitution of another residue at the citrate-binding site (D591V) of human PFK-M resulted in the complete loss of activity. Detailed analyses revealed that the mutated PFK-M subunits formed dimers but were unable to associate into the active tetrameric holoenzyme. These results suggest that stricter control over glycolytic flux developed in metazoans, whose somatic cells are largely characterized by slow proliferation.
format	article
author	Aleksandra Usenik Matic Legiša
author_facet	Aleksandra Usenik Matic Legiša
author_sort	Aleksandra Usenik
title	Evolution of allosteric citrate binding sites on 6-phosphofructo-1-kinase.
title_short	Evolution of allosteric citrate binding sites on 6-phosphofructo-1-kinase.
title_full	Evolution of allosteric citrate binding sites on 6-phosphofructo-1-kinase.
title_fullStr	Evolution of allosteric citrate binding sites on 6-phosphofructo-1-kinase.
title_full_unstemmed	Evolution of allosteric citrate binding sites on 6-phosphofructo-1-kinase.
title_sort	evolution of allosteric citrate binding sites on 6-phosphofructo-1-kinase.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/15018ccb70c943c6ad00ac5e1b28eb4b
work_keys_str_mv	AT aleksandrausenik evolutionofallostericcitratebindingsiteson6phosphofructo1kinase AT maticlegisa evolutionofallostericcitratebindingsiteson6phosphofructo1kinase
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Evolution of allosteric citrate binding sites on 6-phosphofructo-1-kinase.

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