Abstract
Nuclear magnetic resonance (NMR) studies have shown that two distinct folded conformations of staphylococcal nuclease coexist in solution1 and that these two states can interconvert directly without passing through an unfolded state. These experiments have also revealed that the two forms have very different folding kinetics, although the possibility that one component is an obligatory intermediate for the folding of the other form could be discounted1. Here we report NMR data which show that alternative unfolded states are also distinguishable. These observations led us to hypothesize that cis/trans isomerism at a single peptide bond between a proline and its preceding residue might be the origin of the conformational multiplicity. Proline 117 was identified as a likely candidate for the site concerned and a mutant protein, in which Pro 117 was replaced by Gly, was constructed in order to test this. Alternative conformations are not observed in the spectrum of this mutant, lending powerful support to this hypothesis.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Fox, R. O., Evans, P. A. & Dobson, C. M. Nature 320, 192–194 (1986).
Dobson, C. M. & Evans, P. A. Biochemistry 23, 4267–4270 (1984).
Brandts, J. F., Halvorson, H. R. & Brennan, M. Biochemistry 14, 4953–4963 (1975).
Kim, P. S. & Baldwin, R. L. A. Rev. Biochem. 51, 459–489 (1982).
Arnone, A. et al. J. biol. Chem. 246, 2302–2316 (1971).
Dobson, C. M., Evans, P. A. & Williamson, K. L. FEBS Lett. 168, 331–334 (1984).
Lin, L. N. & Brandts, J. F. Biochemistry 23, 5713–5723 (1984).
Schmid, F. X. & Baldwin, R. L. Proc. natn. Acad. Sci. U.S.A. 75, 4764–4768 (1978).
Schmid, F. X., Grafl, R., Wrba, A. & Beintema, J. J. Proc. natn. Acad. Sci. U.S.A. 83, 872–876 (1986).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Evans, P., Dobson, C., Kautz, R. et al. Proline isomerism in staphylococcal nuclease characterized by NMR and site-directed mutagenesis. Nature 329, 266–268 (1987). https://doi.org/10.1038/329266a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/329266a0
This article is cited by
-
Predicting the Solubility of Recombinant Proteins in Escherichia coli
Nature Biotechnology (1991)
-
Identification of a new site of conformational heterogeneity in unfolded ribonuclease A
Journal of Protein Chemistry (1990)
-
Production of Soluble Recombinant Proteins in Bacteria
Nature Biotechnology (1989)
-
Protein Folding: New Twists
Nature Biotechnology (1988)
-
Folding into the right shape
Nature (1987)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.