Optimization of sample preparation for peptide sequencing by MALDI-TOF photofragment mass spectrometry

J M Hettick; D L McCurdy; D C Barbacci; D H Russell

doi:10.1021/ac0102157

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Optimization of sample preparation for peptide sequencing by MALDI-TOF photofragment mass spectrometry

Journal article

Peer reviewed

Optimization of sample preparation for peptide sequencing by MALDI-TOF photofragment mass spectrometry

J M Hettick, D L McCurdy, D C Barbacci and D H Russell

Analytical chemistry (Washington), Vol.73(22), pp.5378-5386

11/15/2001

DOI: 10.1021/ac0102157

PMID: 11816563

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Abstract

This paper describes the optimization of sample preparation for MALDI 193-nm photofragment ion time-of-flight mass spectrometry to sequence small to medium-sized peptides from peptide mixtures. We show that matrix additives, such as fructose and phenylbutyric acid have a dramatic effect on the abundance of fragment ions observed in the post-source decay spectra. A dried-droplet MALDI matrix consisting of 1:1 alpha-cyano-4-hydroxycinnamic acid/fructose proves to be an excellent matrix for photodissociation because [M + H]+ ions are formed with low internal energies, and the photofragment ion spectrum contains high abundances of sequence-informative ions. The addition of fructose appears to improve overall sample homogeneity and durability, as compared to conventional alpha-cyano-4-hydroxycinnamic acid dried-droplet preparations. MALDI-TOF photodissociation is then used to selectively sequence the peptides bradykinin (RPPGFSPFR), des-Arg9 bradykinin (RPPGFSPF), and substance P-amide (RPKPQQFFGLM-NH2) from a mixture of five peptides.

Sequence Analysis, Protein

Specimen Handling

Animals

Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - standards

Humans

Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods

Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - instrumentation

Oligopeptides - chemistry

Coumaric Acids

Fructose

Details

Title: Subtitle: Optimization of sample preparation for peptide sequencing by MALDI-TOF photofragment mass spectrometry
Creators: J M Hettick - Department of Chemistry, Texas A&M University, College Station 77842-3012, USA
D L McCurdy
D C Barbacci
D H Russell
Resource Type: Journal article
Publication Details: Analytical chemistry (Washington), Vol.73(22), pp.5378-5386
DOI: 10.1021/ac0102157
PMID: 11816563
NLM abbreviation: Anal Chem
ISSN: 0003-2700
eISSN: 1520-6882
Publisher: United States
Grant note: P30-ESO9106 / NIEHS NIH HHS
Language: English
Date published: 11/15/2001
Academic Unit: Chemistry
Record Identifier: 9983985925502771

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