Microwave-assisted specific chemical digestion for rapid protein identification

Lin Hua, Low Teck Yew, Siu Kwan Sze

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)586-591
Number of pages6
JournalProteomics
Volume6
Issue number2
DOIs
Publication statusPublished - 1 Jan 2006
Externally publishedYes

Fingerprint

Microwaves
formic acid
Proteolysis
Myoglobin
Proteins
Escherichia coli K12
Peptides
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Microwave irradiation
Hydrolysis
Escherichia coli
Gels
Purification
Acids
Irradiation

Keywords

  • Formic acid
  • MALDI
  • Microwave-assisted digestion
  • Protein identification
  • TOF MS

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Microwave-assisted specific chemical digestion for rapid protein identification. / Hua, Lin; Teck Yew, Low; Sze, Siu Kwan.

In: Proteomics, Vol. 6, No. 2, 01.01.2006, p. 586-591.

Research output: Contribution to journalArticle

Hua, Lin ; Teck Yew, Low ; Sze, Siu Kwan. / Microwave-assisted specific chemical digestion for rapid protein identification. In: Proteomics. 2006 ; Vol. 6, No. 2. pp. 586-591.
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