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Multiple-turnover thio-ATP hydrolase and phospho-enzyme intermediate formation activities catalyzed by an RNA enzyme

Ribozymes that phosphorylate internal 2′-OH positions mimic the first mechanistic step of P-type ATPase enzymes by forming a phospho-enzyme intermediate. We previously described 2′-autophosphorylation and autothiophosphorylation by the 2PTmin3.2 ribozyme. In the present work we demonstrate that the...

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Detalles Bibliográficos
Autores principales: Saran, Dayal, Held, Daniel M., Burke, Donald H.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2006
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1484251/
https://www.ncbi.nlm.nih.gov/pubmed/16790565
http://dx.doi.org/10.1093/nar/gkl413
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author Saran, Dayal
Held, Daniel M.
Burke, Donald H.
author_facet Saran, Dayal
Held, Daniel M.
Burke, Donald H.
author_sort Saran, Dayal
collection PubMed
description Ribozymes that phosphorylate internal 2′-OH positions mimic the first mechanistic step of P-type ATPase enzymes by forming a phospho-enzyme intermediate. We previously described 2′-autophosphorylation and autothiophosphorylation by the 2PTmin3.2 ribozyme. In the present work we demonstrate that the thiophosphorylated form of this ribozyme can de-thiophosphorylate in the absence of ATPγS. Identical ionic conditions yield a thiophosphorylated strand when ATPγS is included, thus effecting a net ATPγS hydrolysis. The de-thiophosphorylation step is nearly independent of pH over the range of 6.3–8.5 and does not require a specifically folded RNA structure, but this step is greatly stimulated by transition metal ions. By monitoring thiophosphate release, we observe 29–46 ATPγS hydrolyzed per ribozyme strand in 24 h, corresponding to a turnover rate of 1.2–2.0 h(−1). The existence of an ATP- (or thio-ATP-)powered catalytic cycle raises the possibility of using ribozymes to transduce chemical energy into mechanical work for nucleic acid nanodevices.
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spelling pubmed-14842512006-07-18 Multiple-turnover thio-ATP hydrolase and phospho-enzyme intermediate formation activities catalyzed by an RNA enzyme Saran, Dayal Held, Daniel M. Burke, Donald H. Nucleic Acids Res Article Ribozymes that phosphorylate internal 2′-OH positions mimic the first mechanistic step of P-type ATPase enzymes by forming a phospho-enzyme intermediate. We previously described 2′-autophosphorylation and autothiophosphorylation by the 2PTmin3.2 ribozyme. In the present work we demonstrate that the thiophosphorylated form of this ribozyme can de-thiophosphorylate in the absence of ATPγS. Identical ionic conditions yield a thiophosphorylated strand when ATPγS is included, thus effecting a net ATPγS hydrolysis. The de-thiophosphorylation step is nearly independent of pH over the range of 6.3–8.5 and does not require a specifically folded RNA structure, but this step is greatly stimulated by transition metal ions. By monitoring thiophosphate release, we observe 29–46 ATPγS hydrolyzed per ribozyme strand in 24 h, corresponding to a turnover rate of 1.2–2.0 h(−1). The existence of an ATP- (or thio-ATP-)powered catalytic cycle raises the possibility of using ribozymes to transduce chemical energy into mechanical work for nucleic acid nanodevices. Oxford University Press 2006 2006-06-21 /pmc/articles/PMC1484251/ /pubmed/16790565 http://dx.doi.org/10.1093/nar/gkl413 Text en © 2006 The Author(s)
spellingShingle Article
Saran, Dayal
Held, Daniel M.
Burke, Donald H.
Multiple-turnover thio-ATP hydrolase and phospho-enzyme intermediate formation activities catalyzed by an RNA enzyme
title Multiple-turnover thio-ATP hydrolase and phospho-enzyme intermediate formation activities catalyzed by an RNA enzyme
title_full Multiple-turnover thio-ATP hydrolase and phospho-enzyme intermediate formation activities catalyzed by an RNA enzyme
title_fullStr Multiple-turnover thio-ATP hydrolase and phospho-enzyme intermediate formation activities catalyzed by an RNA enzyme
title_full_unstemmed Multiple-turnover thio-ATP hydrolase and phospho-enzyme intermediate formation activities catalyzed by an RNA enzyme
title_short Multiple-turnover thio-ATP hydrolase and phospho-enzyme intermediate formation activities catalyzed by an RNA enzyme
title_sort multiple-turnover thio-atp hydrolase and phospho-enzyme intermediate formation activities catalyzed by an rna enzyme
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1484251/
https://www.ncbi.nlm.nih.gov/pubmed/16790565
http://dx.doi.org/10.1093/nar/gkl413
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