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Analysis of a nuclease activity of catalytic domain of Thermus thermophilus MutS2 by high-accuracy mass spectrometry
Electrospray ionization with Fourier-transform ion cyclotron resonance mass spectrometry (ESI–FT ICR MS) is a powerful tool for analyzing the precise structural features of biopolymers, including oligonucleotides. Here, we described the detailed characterization of a newly discovered nuclease activi...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Oxford University Press
2007
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1976465/ https://www.ncbi.nlm.nih.gov/pubmed/17686785 http://dx.doi.org/10.1093/nar/gkm575 |
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author | Fukui, Kenji Takahata, Yoshio Nakagawa, Noriko Kuramitsu, Seiki Masui, Ryoji |
author_facet | Fukui, Kenji Takahata, Yoshio Nakagawa, Noriko Kuramitsu, Seiki Masui, Ryoji |
author_sort | Fukui, Kenji |
collection | PubMed |
description | Electrospray ionization with Fourier-transform ion cyclotron resonance mass spectrometry (ESI–FT ICR MS) is a powerful tool for analyzing the precise structural features of biopolymers, including oligonucleotides. Here, we described the detailed characterization of a newly discovered nuclease activity of the C-terminal domain of Thermus thermophilus MutS2 (ttMutS2). Using this method, the length, nucleotide content and nature of the 5′- and 3′-termini of the product oligonucleotides were accurately identified. It is revealed that the C-terminal domain of ttMutS2 incised the phosphate backbone of oligodeoxynucleotides non-sequence-specifically at the 3′ side of the phosphates. The simultaneous identification of the innumerable fragments was achieved by the extremely high-accuracy of ESI–FT ICR MS. |
format | Text |
id | pubmed-1976465 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2007 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-19764652007-09-26 Analysis of a nuclease activity of catalytic domain of Thermus thermophilus MutS2 by high-accuracy mass spectrometry Fukui, Kenji Takahata, Yoshio Nakagawa, Noriko Kuramitsu, Seiki Masui, Ryoji Nucleic Acids Res Methods Online Electrospray ionization with Fourier-transform ion cyclotron resonance mass spectrometry (ESI–FT ICR MS) is a powerful tool for analyzing the precise structural features of biopolymers, including oligonucleotides. Here, we described the detailed characterization of a newly discovered nuclease activity of the C-terminal domain of Thermus thermophilus MutS2 (ttMutS2). Using this method, the length, nucleotide content and nature of the 5′- and 3′-termini of the product oligonucleotides were accurately identified. It is revealed that the C-terminal domain of ttMutS2 incised the phosphate backbone of oligodeoxynucleotides non-sequence-specifically at the 3′ side of the phosphates. The simultaneous identification of the innumerable fragments was achieved by the extremely high-accuracy of ESI–FT ICR MS. Oxford University Press 2007-08 2007-08-07 /pmc/articles/PMC1976465/ /pubmed/17686785 http://dx.doi.org/10.1093/nar/gkm575 Text en © 2007 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methods Online Fukui, Kenji Takahata, Yoshio Nakagawa, Noriko Kuramitsu, Seiki Masui, Ryoji Analysis of a nuclease activity of catalytic domain of Thermus thermophilus MutS2 by high-accuracy mass spectrometry |
title | Analysis of a nuclease activity of catalytic domain of Thermus thermophilus MutS2 by high-accuracy mass spectrometry |
title_full | Analysis of a nuclease activity of catalytic domain of Thermus thermophilus MutS2 by high-accuracy mass spectrometry |
title_fullStr | Analysis of a nuclease activity of catalytic domain of Thermus thermophilus MutS2 by high-accuracy mass spectrometry |
title_full_unstemmed | Analysis of a nuclease activity of catalytic domain of Thermus thermophilus MutS2 by high-accuracy mass spectrometry |
title_short | Analysis of a nuclease activity of catalytic domain of Thermus thermophilus MutS2 by high-accuracy mass spectrometry |
title_sort | analysis of a nuclease activity of catalytic domain of thermus thermophilus muts2 by high-accuracy mass spectrometry |
topic | Methods Online |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1976465/ https://www.ncbi.nlm.nih.gov/pubmed/17686785 http://dx.doi.org/10.1093/nar/gkm575 |
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