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Microwave‐assisted specific chemical digestion for rapid protein identification
Author(s) -
Hua Lin,
Low Teck Yew,
Sze Siu Kwan
Publication year - 2006
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200500304
Subject(s) - myoglobin , formic acid , chemistry , chromatography , hydrolysis , peptide , peptide mass fingerprinting , biochemistry , mass spectrometry , fragmentation (computing) , proteomics , biology , ecology , gene
We have developed a rapid microwave‐assisted protein digestion technique based on classic acid hydrolysis reaction with 2% formic acid solution. In this mild chemical environment, proteins are hydrolyzed to peptides, which can be directly analyzed by MALDI‐MS or ESI‐MS without prior sample purification. Dilute formic acid cleaves proteins specifically at the C‐terminal of aspartyl (Asp) residues within 10 min of exposure to microwave irradiation. By adjusting the irradiation time, we found that the extent of protein fragmentation can be controlled, as shown by the single fragmentation of myoglobin at the C‐terminal of any of the Asp residues. The efficacy and simplicity of this technique for protein identification are demonstrated by the peptide mass maps of in‐gel digested myoglobin and BSA, as well as proteins isolated from Escherichia coli K12 cells.