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Home / Equine Research Bank / Proc Natl Acad Sci U S A / Semisynthetic cytochrome c.
Proceedings of the National Academy of Sciences of the United States of America1977; 74(10); 4248-4250; doi: 10.1073/pnas.74.10.4248

Semisynthetic cytochrome c.

Abstract: Horse heart cytochrome c can be split with cyanogen bromide into a heme peptide (residues 1-65) and a nonheme peptide (residues 66-104). In a process involving (i) complex formation between the two fragments and (ii) restoration of the severed peptide linkage, a fully active cytochrome c preparation can be re-formed. Use has been made of this process to couple the heme peptide to peptide 66-104 synthesized by the Merrifield solid-phase procedure. The semisynthetic product formed in this manner is indistinguishable from reconstituted cytochrome c prepared with nonsynthetic peptide 66-104.
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Publication Date: 1977-10-01 PubMed ID: 200910PubMed Central: PMC431916DOI: 10.1073/pnas.74.10.4248Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article discusses a method to divide horse heart cytochrome c into two fragments – a heme peptide and a nonheme peptide, then using these two fragments to recreate a fully active cytochrome c molecule. This restored molecule, a semisynthetic product, matched the characteristics of a cytochrome c formed with nonsynthetic peptide 66-104.

Division of Horse Heart Cytochrome c

  • The research begins with the division of horse heart cytochrome c into two distinct parts – a heme peptide, incorporating residues 1-65, and a nonheme peptide, encompassing residues 66-104. This process is accomplished using cyanogen bromide, a substance known for its ability to cleave proteins at methionine residues.

Restoration of the Severed Peptide

  • Following the division process, researchers reorganize the two fragments, the heme peptide and the nonheme peptide. The process entails forming a complex between the two parts and carrying out the restoration of the original peptide linkage that was disrupted during the cleavage.
  • The end product of this process is a completely active cytochrome c molecule, successfully restored from its fragmented pieces.

Use of the Merrifield Solid-Phase Procedure

  • The study additionally explores coupling the heme peptide with a synthesized peptide 66-104, derived via the Merrifield solid-phase procedure. This specific procedure is a widely recognized approach for peptide synthesis. It relies on the step-by-step addition of amino acids while the growing peptide chain remains attached to an insoluble resin, making the purification process straightforward and efficient.

Creation and Validation of The Semisynthetic Product

  • When the heme peptide is combined with the synthesized peptide 66-104, a semisynthetic product is formed. This output is indistinguishable from reconstituted cytochrome c that would be prepared with nonsynthetic peptide 66-104, effectively validating the semi-synthesis method as detailed in this study.

Cite This Article

APA
Barstow LE, Young RS, Yakali E, Sharp JJ, O'Brien JC, Berman PW, Harbury HA. (1977). Semisynthetic cytochrome c. Proc Natl Acad Sci U S A, 74(10), 4248-4250. https://doi.org/10.1073/pnas.74.10.4248

Publication

Proceedings of the National Academy of Sciences of the United States of America
ISSN: 0027-8424
NlmUniqueID: 7505876
Country: United States
Language: English
Volume: 74
Issue: 10
Pages: 4248-4250

Researcher Affiliations

Barstow, L E
    Young, R S
      Yakali, E
        Sharp, J J
          O'Brien, J C
            Berman, P W
              Harbury, H A

                MeSH Terms

                • Amino Acids / analysis
                • Animals
                • Chemical Phenomena
                • Chemistry
                • Cytochrome c Group / analysis
                • Cytochrome c Group / chemical synthesis
                • Horses
                • Myocardium / enzymology
                • Peptides / analysis
                • Peptides / chemical synthesis
                • Radioimmunoassay

                References

                This article includes 12 references
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                Citations

                This article has been cited 2 times.
                1. Wallace CJ, Offord RE. The semisynthesis of fragments corresponding to residues 66-104 of horse heart cytochrome c.. Biochem J 1979 Apr 1;179(1):169-82.
                  doi: 10.1042/bj1790169pubmed: 224859google scholar: lookup
                2. Boon PJ, Tesser GI, Nivard RJ. Semisynthetic horse heart [65-homoserine]cytochrome c from three fragments.. Proc Natl Acad Sci U S A 1979 Jan;76(1):61-5.
                  doi: 10.1073/pnas.76.1.61pubmed: 218205google scholar: lookup
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