The design and production of semisynthetic ribonucleases with increased thermostability by incorporation of S‐peptide analogues with enhanced helical stability

Colin Mitchinson; Robert L. Baldwin

doi:10.1002/prot.340010106

journal article Jan 01, 1986

The design and production of semisynthetic ribonucleases with increased thermostability by incorporation of S‐peptide analogues with enhanced helical stability

Colin Mitchinson Robert L. Baldwin

Proteins: Structure, Function, and Bioinformatics Vol. 1 No. 1 pp. 23-33 · Wiley

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Abstract

AbstractRecent work has shown that, with synthetic analogues of C‐peptide (residues 1–13 of ribonuclease A), the stability of the peptide helix in H2O depends strongly on the charge on the N‐terminal residue. We have asked whether, in semisynthetic ribonuclease S reconstituted from S‐protein plus an analogue of S‐peptide (1–15), the stability of the peptide helix is correlated with the Tm of the reconstituted ribonuclease S. Six peptides have been made, which contain Glu9 → Leu, a blocked α‐COO− group (CONH2), and either Gln11 or Glu11. The N‐terminal residue has been varied; its charge varies from +2 (Lys) to −1 (succinyl‐Ala). We have measured the stability of the peptide helix, the affinity of the peptide for S‐protein (by C.D. titration), and the thermal stability of the reconstituted ribonuclease S.All six peptide analogues show strongly enhanced helix formation compared to either S‐peptide (1–15) or (1–19), and the helix content increases as the charge on the N‐terminal residue changes from +2 to −1. All six peptides show increased affinity for S‐protein compared to S‐peptide (1–19), and all six reconstituted ribonucleases S show an increase in Tm compared to the protein with S‐peptide (1–19). The Tm increases as the charge on residue 1 changes from +2 to −1. The largest increment in Tm is 6°.The results suggest that the stability of a protein can be increased by enhancing the stability of its secondary structure.

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Citations

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References

Details

Published: Jan 01, 1986
Vol/Issue: 1(1)
Pages: 23-33
License: View

Authors

National Institute of Chemistry, Hajdrihova 19, Ljubljana SI 1115, Slovenia; and Department of Biochemistry, Beckman Center, Stanford University Medical Center, Stanford, CA 94305-5307

Cite This Article

Colin Mitchinson, Robert L. Baldwin (1986). The design and production of semisynthetic ribonucleases with increased thermostability by incorporation of S‐peptide analogues with enhanced helical stability. Proteins: Structure, Function, and Bioinformatics, 1(1), 23-33. https://doi.org/10.1002/prot.340010106

The design and production of semisynthetic ribonucleases with increased thermostability by incorporation of S‐peptide analogues with enhanced helical stability

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