Investigation of pyrophosphate versus ATP substrate selection in the Entamoeba histolytica acetate kinase

Abstract Acetate kinase (ACK; E.C. 2.7.2.1), which catalyzes the interconversion of acetate and acetyl phosphate, is nearly ubiquitous in bacteria but is present only in one genus of archaea and certain eukaryotic microbes. All ACKs utilize ATP/ADP as the phosphoryl donor/acceptor in the respective...

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Autores principales:	Thanh Dang, Cheryl Ingram-Smith
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/51c7068af56a439080e11fc384802620
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spelling	oai:doaj.org-article:51c7068af56a439080e11fc3848026202021-12-02T12:32:19ZInvestigation of pyrophosphate versus ATP substrate selection in the Entamoeba histolytica acetate kinase10.1038/s41598-017-06156-52045-2322https://doaj.org/article/51c7068af56a439080e11fc3848026202017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-06156-5https://doaj.org/toc/2045-2322Abstract Acetate kinase (ACK; E.C. 2.7.2.1), which catalyzes the interconversion of acetate and acetyl phosphate, is nearly ubiquitous in bacteria but is present only in one genus of archaea and certain eukaryotic microbes. All ACKs utilize ATP/ADP as the phosphoryl donor/acceptor in the respective directions of the reaction (acetate + ATP $${\boldsymbol{\leftrightarrows }}$$ ⇆ acetyl phosphate + ADP), with the exception of the Entamoeba histolytica ACK (EhACK) which uses pyrophosphate (PPi)/inorganic phosphate (Pi) (acetyl phosphate + Pi $${\boldsymbol{\leftrightarrows }}$$ ⇆ acetate + PPi). Structural analysis and modeling of EhACK indicated steric hindrance by active site residues constricts entry to the adenosine pocket as compared to ATP-utilizing Methanosarcina thermophila ACK (MtACK). Reciprocal alterations were made to enlarge the adenosine pocket of EhACK and reduce that of MtACK. The EhACK variants showed a step-wise increase in ADP and ATP binding but were still unable to use these as substrates, and enzymatic activity with Pi/PPi was negatively impacted. Consistent with this, ATP utilization by MtACK variants was negatively affected but the alterations were not sufficient to convert this enzyme to Pi/PPi utilization. Our results suggest that controlling access to the adenosine pocket can contribute to substrate specificity but is not the sole determinant.Thanh DangCheryl Ingram-SmithNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Thanh Dang Cheryl Ingram-Smith Investigation of pyrophosphate versus ATP substrate selection in the Entamoeba histolytica acetate kinase
description	Abstract Acetate kinase (ACK; E.C. 2.7.2.1), which catalyzes the interconversion of acetate and acetyl phosphate, is nearly ubiquitous in bacteria but is present only in one genus of archaea and certain eukaryotic microbes. All ACKs utilize ATP/ADP as the phosphoryl donor/acceptor in the respective directions of the reaction (acetate + ATP $${\boldsymbol{\leftrightarrows }}$$ ⇆ acetyl phosphate + ADP), with the exception of the Entamoeba histolytica ACK (EhACK) which uses pyrophosphate (PPi)/inorganic phosphate (Pi) (acetyl phosphate + Pi $${\boldsymbol{\leftrightarrows }}$$ ⇆ acetate + PPi). Structural analysis and modeling of EhACK indicated steric hindrance by active site residues constricts entry to the adenosine pocket as compared to ATP-utilizing Methanosarcina thermophila ACK (MtACK). Reciprocal alterations were made to enlarge the adenosine pocket of EhACK and reduce that of MtACK. The EhACK variants showed a step-wise increase in ADP and ATP binding but were still unable to use these as substrates, and enzymatic activity with Pi/PPi was negatively impacted. Consistent with this, ATP utilization by MtACK variants was negatively affected but the alterations were not sufficient to convert this enzyme to Pi/PPi utilization. Our results suggest that controlling access to the adenosine pocket can contribute to substrate specificity but is not the sole determinant.
format	article
author	Thanh Dang Cheryl Ingram-Smith
author_facet	Thanh Dang Cheryl Ingram-Smith
author_sort	Thanh Dang
title	Investigation of pyrophosphate versus ATP substrate selection in the Entamoeba histolytica acetate kinase
title_short	Investigation of pyrophosphate versus ATP substrate selection in the Entamoeba histolytica acetate kinase
title_full	Investigation of pyrophosphate versus ATP substrate selection in the Entamoeba histolytica acetate kinase
title_fullStr	Investigation of pyrophosphate versus ATP substrate selection in the Entamoeba histolytica acetate kinase
title_full_unstemmed	Investigation of pyrophosphate versus ATP substrate selection in the Entamoeba histolytica acetate kinase
title_sort	investigation of pyrophosphate versus atp substrate selection in the entamoeba histolytica acetate kinase
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/51c7068af56a439080e11fc384802620
work_keys_str_mv	AT thanhdang investigationofpyrophosphateversusatpsubstrateselectionintheentamoebahistolyticaacetatekinase AT cherylingramsmith investigationofpyrophosphateversusatpsubstrateselectionintheentamoebahistolyticaacetatekinase
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Investigation of pyrophosphate versus ATP substrate selection in the Entamoeba histolytica acetate kinase

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