Specific cleavage of amino side chains of serine and threonine in peptides and proteins with S-ethyltrifluorothioacetate vapor.
A vapor of S-ethyltrifluorothioacetate was found to specifically cleave the amino side of serine and threonine peptide bonds. The cleavage reactions were carried out at 50 degrees C for 6 h-24 h or at 30 degrees C for 24 h. When vapors were generated in a solution containing several conventional organic solvents, the cleavage reactions were reduced or stopped, or modification took place. When the reagent vapor was made in an aqueous solution, the cleavage reaction at glycine residues was enhanced. This reagent did not oxidize any amino acid residues, such as methionine, cysteine and tryptophan. The cleavage was also effective on proteins on membranes blotted or electroblotted from polyacrylamide gels. This method therefore may be used for the peptide mass fingerprinting [Patterson, S. D. (1995) Electrophoresis 16, 1104-1114] after two-dimensional electrophoresis.
Kamo
M
M
Research Institute for Biosciences, Science University of Tokyo, Yamazaki, Chiba, Japan.
Tsugita
A
A
D000426:Alcohol Dehydrogenase / Q000737:chemistry; D000595:Amino Acid Sequence; D002213:Capsid / Q000737:chemistry; D005463:Fluoroacetates; D005740:Gases; D005934:Glucagon / Q000737:chemistry; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000737:chemistry; D009037:Motilin / Q000737:chemistry; D009211:Myoglobin / Q000737:chemistry; D010047:Ovalbumin / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry; D010449:Peptide Mapping / Q000379:methods*; D010455:Peptides / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D017421:Sequence Analysis; D012694:Serine / Q000737:chemistry*; D016339:Spectrometry, Mass, Fast Atom Bombardment; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013912:Threonine / Q000737:chemistry*; D014269:Trifluoroacetic Acid / Q000737:chemistry; D018822:alpha-Endorphin / Q000737:chemistry
D005463:Fluoroacetates; D005740:Gases; D009211:Myoglobin; D010446:Peptide Fragments; D010455:Peptides; D011506:Proteins; D013912:Threonine; C028085:ethylthiol trifluoroacetate; D012694:Serine; D009037:Motilin; D018822:alpha-Endorphin; D010047:Ovalbumin; D005934:Glucagon; D014269:Trifluoroacetic Acid; D000426:Alcohol Dehydrogenase