A method for identifying the carboxy terminal amino acid of a protein.
A highly sensitive new method for identifying the carboxy terminus of a protein was developed. The carboxyl terminal amino acid was racemized by reaction with acetic anhydride. The resulting modified protein was subjected to acid hydrolysis. The hydrolysate was derivatized with (+)-1-(9-fluorenyl)ethyl chloroformate to give fluorescent amino acid diastereomers. The amino acid diastereomers were separated on a reversed-phase column. Only carboxyl terminal amino acids give a D-amino acid. Application of this method was described for the isolation and identification of carboxyl terminal peptides from an enzymatic digest of a protein.
Hayashi
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Toray Research Center, Inc., Kamakura, Japan.
Sasagawa
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D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis*; D002623:Chemistry Techniques, Analytical / Q000379:methods; D002845:Chromatography / Q000379:methods; D005449:Fluorenes; D005453:Fluorescence; D013006:Growth Hormone / Q000032:analysis; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification; D011506:Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D014357:Trypsin / Q000378:metabolism
D000596:Amino Acids; D005449:Fluorenes; D010455:Peptides; D011506:Proteins; D013006:Growth Hormone; C054007:1-(9-fluorenyl)methyl chloroformate; D014357:Trypsin