Chemical oxidation of tryptic digests to improve sequence coverage in peptide mass fingerprint protein identification

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Title: Chemical oxidation of tryptic digests to improve sequence coverage in peptide mass fingerprint protein identification
Author: Lucas, Jessica Elaine
Abstract: Peptide mass fingerprinting (PMF ) of protein digests is a widely -accepted method for protein identification in MS -based proteomic studies . Matrix -assisted laser desorption /ionization mass spectrometry (MALDI ) is the technique of choice in PMF experiments . The success of protein identification in a PMF experiment is directly related to the amount of amino acid sequence coverage . In an effort to increase the amount of sequence information obtained in a MALDI PMF experiment , performic acid oxidation is performed on tryptic digests of known proteins . Performic acid was chosen as the chemical oxidant due to the ease of use and to the selective oxidation of cysteine , methionine , and tryptophan residues . In experiments performed in our laboratory , performic acid oxidation either increased or did not affect protein sequence coverage in PMF experiments when oxidized tryptic digests were analyzed by MALDI . Negative mode MALDI data were acquired , as well as positive mode MALDI data , due to the enhanced ionization of cysteic acid -containing peptides in negative mode . Furthermore , the confidence in a protein match is increased by observation of mass shifts indicative of cysteine , methionine , and /or tryptophan in oxidized peptide ion signals when comparing MALDI spectra prior to performic acid oxidation and after oxidation due to the low abundance of these residues in the majority of all known and hypothetical proteins .
URI: http : / /hdl .handle .net /1969 .1 /66
Date: 2004-09-30

Citation

Chemical oxidation of tryptic digests to improve sequence coverage in peptide mass fingerprint protein identification. Available electronically from http : / /hdl .handle .net /1969 .1 /66 .

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