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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...
Autores principales: | , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2006
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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. |
format | Text |
id | pubmed-1484251 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2006 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
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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