CLOSTRIPAIN‐CATALYZED RE‐FORMATION OF A PEPTIDE BOND IN A CYTOCHROME C FRAGMENT COMPLEX*
- 1 October 1981
- journal article
- research article
- Published by Wiley in International Journal of Peptide and Protein Research
- Vol. 18 (4) , 335-342
- https://doi.org/10.1111/j.1399-3011.1981.tb02990.x
Abstract
Enzymatically-catalyzed condensation of cytochrome c fragments, ferrous heme fragment (1-38) and apofragment (39-104), has allowed the back-conversion of cytochrome c complex to native cytochrome c. The conversion was accomplished in 90% (vol/vol) glycerol, a solvent which has been shown to decrease the ionization of the terminal .alpha.-carboxyl group liberated during hydrolysis of a peptide bond. The effect on the pK is probably the main reason the thermodynamic obstacle to re-synthesis is minimized. A 30% conversion to cytochrome c was obtained. The cytochrome c product was distinguished from the noncovalent complex and separated fragments by MW analysis with sodium dodecyl sulfate polyacrylamide gel electrophoresis, by elution from Sephadex G-50 and sulfopropyl-Sephadex in the presence of denaturant, by amino acid analysis of the product purified under complex-dissociation conditions and by spectral analysis of the absorption bands of the heme. This method provides an opportunity to study the covalent rather than the complex form of cytochrome c analogs.Keywords
This publication has 27 references indexed in Scilit:
- ENZYME-CATALYZED FORMATION OF SEMISYNTHETIC STAPHYLOCOCCAL NUCLEASE USING A NEW SYNTHETIC FRAGMENT, [48-GLYCINE] SYNTHETIC-(6-49)International Journal of Peptide and Protein Research, 2009
- Semi-synthetic analogs of cytochrome c at positions 67 and 74Biochemical and Biophysical Research Communications, 1979
- Semisynthetic analogs of cytochrome c reconstructed from natural and synthetic peptidesBiochimica et Biophysica Acta (BBA) - Protein Structure, 1979
- Enzymic resynthesis of the hydrolyzed peptide bond(s) in ribonuclease SBiochemistry, 1979
- Synthesis of peptide bonds by proteinases. Addition of organic cosolvents shifts peptide bond equilibriums toward synthesisBiochemistry, 1978
- The cytochrome fold and the evolution of bacterial energy metabolismJournal of Molecular Biology, 1976
- Reconstitution of horse heart cytochrome c: Reformation of the peptide bond linking residues 65 and 66Biochemical and Biophysical Research Communications, 1974
- S-peptide-S-protein system. Model for hormone-receptor interactionAccounts of Chemical Research, 1973
- Electrophoretic analysis of the major polypeptides of the human erythrocyte membraneBiochemistry, 1971
- The insensitivity of the 695 nm band of horse heart ferricytochrome
c to protein conformationBiochemical and Biophysical Research Communications, 1971