Evaluation of microwave-accelerated residue-specific acid cleavage for proteomic applications.
Microwave-accelerated proteolysis using acetic acid has been shown to occur specifically on either or both sides of aspartic acid residues. This chemical cleavage has been applied to ovalbumin and several model peptides to test the effect on some of the more common post-translational modifications. No oxidation of methionine or cysteine was observed; however, hydrolysis of phosphate groups proceeds at a detectable rate. Acid cleavage was also extended to the yeast ribosome model proteome, where it provided information on 74% of that proteome. Aspartic acid occurs across the proteome with approximately half the frequency of the combined occurrence of the trypsin residues lysine and arginine, and implications of this are considered.
Swatkoski
Stephen
S
Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA.
Gutierrez
Peter
P
Wynne
Colin
C
Petrov
Alexey
A
Dinman
Jonathan D
JD
Edwards
Nathan
N
Fenselau
Catherine
C
D019342:Acetic Acid; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism*; D006868:Hydrolysis; D008872:Microwaves; D008969:Molecular Sequence Data; D010047:Ovalbumin / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D012269:Ribosomal Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
D000596:Amino Acids; D010455:Peptides; D020543:Proteome; D012269:Ribosomal Proteins; D010047:Ovalbumin; D019342:Acetic Acid