10-5+Chapter3++蛋白质化学(F)幻灯片.ppt

Analysis of aa sequence in the polypeptide  (protein sequencing) Sanger.1953, insulin Nobel prize Major step: purification, disulfide bond break(single chain) HCl degradation(6N, 110,180C,36h), aa composition, aa seperation by ion exchange chromatography. N-terminal(DNP, Dansyl-Cl), C-terminal (carboxylase) chemical method, enzymetic method Peptide chain peptide fragment Enzyme. Trypsin (Lys. Arg), Chymotrypsin(Phe, Tyr, Trp), BrCN(Met) Seperation Sequencing aa of peptide fragment Edman reation (degradation, phenlythisoctanate, 异