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Enhanced MALDI-TOF MS Analysis of Phosphopeptides Using an Optimized DHAP/DAHC Matrix

Selecting an appropriate matrix solution is one of the most effective means of increasing the ionization efficiency of phosphopeptides in matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In this study, we systematically assessed matrix combinations of 2, 6...

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Detalles Bibliográficos
Autores principales: Hou, Junjie, Xie, Zhensheng, Xue, Peng, Cui, Ziyou, Chen, Xiulan, Li, Jing, Cai, Tanxi, Wu, Peng, Yang, Fuquan
Formato: Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2842900/
https://www.ncbi.nlm.nih.gov/pubmed/20339515
http://dx.doi.org/10.1155/2010/759690
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author Hou, Junjie
Xie, Zhensheng
Xue, Peng
Cui, Ziyou
Chen, Xiulan
Li, Jing
Cai, Tanxi
Wu, Peng
Yang, Fuquan
author_facet Hou, Junjie
Xie, Zhensheng
Xue, Peng
Cui, Ziyou
Chen, Xiulan
Li, Jing
Cai, Tanxi
Wu, Peng
Yang, Fuquan
author_sort Hou, Junjie
collection PubMed
description Selecting an appropriate matrix solution is one of the most effective means of increasing the ionization efficiency of phosphopeptides in matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In this study, we systematically assessed matrix combinations of 2, 6-dihydroxyacetophenone (DHAP) and diammonium hydrogen citrate (DAHC), and demonstrated that the low ratio DHAP/DAHC matrix was more effective in enhancing the ionization of phosphopeptides. Low femtomole level of phosphopeptides from the tryptic digests of α-casein and β-casein was readily detected by MALDI-TOF-MS in both positive and negative ion mode without desalination or phosphopeptide enrichment. Compared with the DHB/PA matrix, the optimized DHAP/DAHC matrix yielded superior sample homogeneity and higher phosphopeptide measurement sensitivity, particularly when multiple phosphorylated peptides were assessed. Finally, the DHAP/DAHC matrix was applied to identify phosphorylation sites from α-casein and β-casein and to characterize two phosphorylation sites from the human histone H1 treated with Cyclin-Dependent Kinase-1 (CDK1) by MALDI-TOF/TOF MS.
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spelling pubmed-28429002010-03-25 Enhanced MALDI-TOF MS Analysis of Phosphopeptides Using an Optimized DHAP/DAHC Matrix Hou, Junjie Xie, Zhensheng Xue, Peng Cui, Ziyou Chen, Xiulan Li, Jing Cai, Tanxi Wu, Peng Yang, Fuquan J Biomed Biotechnol Methodology Report Selecting an appropriate matrix solution is one of the most effective means of increasing the ionization efficiency of phosphopeptides in matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In this study, we systematically assessed matrix combinations of 2, 6-dihydroxyacetophenone (DHAP) and diammonium hydrogen citrate (DAHC), and demonstrated that the low ratio DHAP/DAHC matrix was more effective in enhancing the ionization of phosphopeptides. Low femtomole level of phosphopeptides from the tryptic digests of α-casein and β-casein was readily detected by MALDI-TOF-MS in both positive and negative ion mode without desalination or phosphopeptide enrichment. Compared with the DHB/PA matrix, the optimized DHAP/DAHC matrix yielded superior sample homogeneity and higher phosphopeptide measurement sensitivity, particularly when multiple phosphorylated peptides were assessed. Finally, the DHAP/DAHC matrix was applied to identify phosphorylation sites from α-casein and β-casein and to characterize two phosphorylation sites from the human histone H1 treated with Cyclin-Dependent Kinase-1 (CDK1) by MALDI-TOF/TOF MS. Hindawi Publishing Corporation 2010 2010-03-21 /pmc/articles/PMC2842900/ /pubmed/20339515 http://dx.doi.org/10.1155/2010/759690 Text en Copyright © 2010 Junjie Hou et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology Report
Hou, Junjie
Xie, Zhensheng
Xue, Peng
Cui, Ziyou
Chen, Xiulan
Li, Jing
Cai, Tanxi
Wu, Peng
Yang, Fuquan
Enhanced MALDI-TOF MS Analysis of Phosphopeptides Using an Optimized DHAP/DAHC Matrix
title Enhanced MALDI-TOF MS Analysis of Phosphopeptides Using an Optimized DHAP/DAHC Matrix
title_full Enhanced MALDI-TOF MS Analysis of Phosphopeptides Using an Optimized DHAP/DAHC Matrix
title_fullStr Enhanced MALDI-TOF MS Analysis of Phosphopeptides Using an Optimized DHAP/DAHC Matrix
title_full_unstemmed Enhanced MALDI-TOF MS Analysis of Phosphopeptides Using an Optimized DHAP/DAHC Matrix
title_short Enhanced MALDI-TOF MS Analysis of Phosphopeptides Using an Optimized DHAP/DAHC Matrix
title_sort enhanced maldi-tof ms analysis of phosphopeptides using an optimized dhap/dahc matrix
topic Methodology Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2842900/
https://www.ncbi.nlm.nih.gov/pubmed/20339515
http://dx.doi.org/10.1155/2010/759690
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