Total synthesis of horse heart cytochrome c. Preparation and characterization of the (1-66)apofragment.
A peptide corresponding to the native (1-66) sequence of horse heart cytochrome c has been synthesized by stepwise automated solid-phase methods on PAM resin. The course of the synthesis has been monitored by several analytical methods including quantitative ninhydrin and Edman degradation. After HF cleavage, the peptide has been purified by a combination of semipreparative ion-exchange and RP-HPLC. The homogeneity of the purified synthetic peptide has been determined by different criteria including HPLC, amino-acid composition, electrophoresis, antibody binding, tryptic and chymotryptic peptide mapping. After deprotection of the Acm-Cys residues and CNBr cleavage of the Met65-Glu66 peptide bond with simultaneous transformation of the Met65 residue into the activated C-terminal [Hse65]lactone, this purified synthetic peptide has been utilized for conformation-assisted joining experiments in combination with synthetic (66-104) to produce fully synthetic [Hse65]apocytochrome c. This latter, after mitochondria-mediated stereospecific heme insertion, has given a functional molecule corresponding to native horse heart holocytochrome c.
Di Bello
C
C
Institute of Industrial Chemistry, University of Padua, Italy.
Gozzini
L
L
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001059:Apoproteins / Q000138:chemical synthesis* / Q000187:drug effects / Q000276:immunology; D002918:Chymotrypsin / Q000494:pharmacology; D003574:Cytochrome c Group / Q000138:chemical synthesis* / Q000187:drug effects / Q000276:immunology; D006736:Horses; D008969:Molecular Sequence Data; D009206:Myocardium / Q000737:chemistry; D010446:Peptide Fragments / Q000138:chemical synthesis; D014357:Trypsin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D001059:Apoproteins; D003574:Cytochrome c Group; D010446:Peptide Fragments; D002918:Chymotrypsin; D014357:Trypsin