Document Detail

A strategy to improve peptide mass fingerprinting matches through the optimization of matrix-assisted laser desorption/ionization matrix selection and formulation.
MedLine Citation:
PMID:  14760719     Owner:  NLM     Status:  MEDLINE    
Peptide mass fingerprinting (PMF) is a powerful technique in which experimentally measured m/z values of peptides that result from a protein digest form the basis for a characteristic fingerprint of the intact protein. Due to its propensity to generate singly-charged ions, along with its relative insensitivity to salts and buffers, matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) is the MS method of choice for PMF. The qualitative features of a MALDI-MS mass spectrum can be selectively tuned by varying the matrix and the solvent system used to prepare the matrix. The selective tuning of MALDI-MS mass spectra in order to optimize PMF results is addressed in this paper. Carbonic anhydrase, hemoglobin alpha- and beta-chain, and myoglobin were digested with trypsin, and the digest was analyzed with MALDI-MS. 2,5-Dihydroxybenzoic acid (2,5-DHB) and alpha-cyano-4-hydroxycinnamic acid were prepared, using five different solvent systems: (A) 99% acetone; (B) 50% acetonitrile (ACN), 0.1% trifluoroacetic acid (TFA); (C) 75% ACN, 0.1% TFA; (D) formic acid:H(2)O: 2-propanol (1:3:2); and (E) H(2)O:MeOH (2:1). Each protein was found to have a different optimum solvent system for PMF. Generally, better PMF results were obtained with 2,5-DHB. The best PMF results were obtained when all of the mass spectral data for a particular protein digest were convolved.
Neerav D Padliya; Troy D Wood
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Proteomics     Volume:  4     ISSN:  1615-9853     ISO Abbreviation:  Proteomics     Publication Date:  2004 Feb 
Date Detail:
Created Date:  2004-02-04     Completed Date:  2004-09-21     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  101092707     Medline TA:  Proteomics     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  466-73     Citation Subset:  IM    
Department of Chemistry, University at Buffalo, The State University of New York, Buffalo, NY, USA.
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MeSH Terms
Amino Acid Sequence
Carbonic Anhydrases / chemistry*
Databases, Protein
Hemoglobins / chemistry*
Molecular Sequence Data
Myoglobin / chemistry*
Peptide Mapping
Peptides / chemistry
Solvents / chemistry
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
Reg. No./Substance:
0/Hemoglobins; 0/Myoglobin; 0/Peptides; 0/Solvents; EC Anhydrases

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