High-throughput method for on-target performic acid oxidation of MALDI-deposited samples.
An information-rich on-target performic acid oxidation method, which is compatible with alkylation for differentiation of free cysteine versus disulfide-containing peptides, is described. On-target oxidation is achieved using performic acid vapor to oxidize disulfide-containing peptides and/or small proteins on the matrix-assisted laser desorption/ionization (MALDI) sample deposits. The on-target oxidation method is preferred over solution-phase oxidation methods because (1) less sample handling is required, (2) oxidation throughput is drastically increased and (3) ion suppression effects are reduced because performic acid is not added directly to the MALDI spot. The utility of this method is demonstrated by simultaneous oxidation of multiple MALDI sample deposits containing model disulfide-linked peptides, intact bovine insulin and a bovine ribonuclease A proteolytic digest.
Williams
Brad J
BJ
Department of Chemistry, Texas A&M University, College Station, TX 77843, USA.
Russell
William K
WK
Russell
David H
DH
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D004220:Disulfides / Q000737:chemistry; D005561:Formates / Q000737:chemistry*; D057166:High-Throughput Screening Assays / Q000191:economics / Q000379:methods; D006728:Hormones / Q000737:chemistry; D007328:Insulin / Q000737:chemistry; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011506:Proteins / Q000737:chemistry*; D012259:Ribonuclease, Pancreatic / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000191:economics / Q000379:methods*
D004220:Disulfides; D005561:Formates; D006728:Hormones; D007328:Insulin; D011506:Proteins; C008817:peroxyformic acid; D012259:Ribonuclease, Pancreatic