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Guanidinium Chloride Induction of Partial Unfolding in Amide Proton Exchange in RNase A
Abstract
Amide (NH) proton exchange rates were measured in 0.0 to 0.7 M guanidinium chloride (GdmCl) for 23 slowly exchanging peptide NH protons of ribonuclease A (RNase A) at pH * 5.5 (uncorrected pH measured in D 2 O), 34°C. The purpose was to find out whether GdmCl induces exchange through binding to exchange intermediates that are partly or wholly unfolded. It was predicted that, when the logarithm of the exchange rate is plotted as a function of the molarity of GdmCl, the slope should be a measure of the amount of buried surface area exposed to GdmCl in the exchange intermediate. The results indicate that these concentrations of GdmCl do induce exchange by means of a partial unfolding mechanism for all 23 protons; this implies that exchange reactions can be used to study the unfolding and stability of local regions. Of the 23 protons, nine also show a second mechanism of exchange at lower concentrations of GdmCl, a mechanism that is nearly independent of GdmCl concentration and is termed "limited structural fluctuation."
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Cited by
Protein Folding Intermediates: Native-State Hydrogen Exchange
Bai, Y., Sosnick, T. R., Mayne, L., & Englander, S. W. (1995) · Science · doi:10.1126/science.7618079 · Cited by 990
Mechanisms and uses of hydrogen exchange
Englander, S. W., Sosnick, T. R., Englander, J. J., & Mayne, L. (1996) · Current Opinion in Structural Biology · doi:10.1016/s0959-440x(96)80090-x · Cited by 383
Monera, O. D., Kay, C. M., & Hodges, R. S. (1994) · Protein Science · doi:10.1002/pro.5560031110 · Cited by 351
Detection of rare partially folded molecules in equilibrium with the native conformation of RNaseH
Chamberlain, A. K., Handel, T. M., & Marqusee, S. (1996) · Nature Structural & Molecular Biology · doi:10.1038/nsb0996-782 · Cited by 318
Protein stability parameters measured by hydrogen exchange
Bai, Y., Milne, J. S., Mayne, L., & Englander, S. W. (1994) · Proteins: Structure, Function, and Bioinformatics · doi:10.1002/prot.340200103 · Cited by 307
Metadata
- Authors
- Mayo, S. L., & Baldwin, R. L.
- Venue
- Science
- Published
- 1993-11-05
- Volume
- 262
- Issue
- 5135
- Pages
- 873-876
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- OpenAlex
- W2049709989
- Cited by
- 180
- References
- 0
- FWCI
- 3.25
- Type
- journal article
- Language
- en
- Open access
- closed
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