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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...

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Detalles Bibliográficos
Autores principales: Fukui, Kenji, Takahata, Yoshio, Nakagawa, Noriko, Kuramitsu, Seiki, Masui, Ryoji
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2007
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.
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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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