The Rate of Isomerisation of Peptidyl-proline Bonds as a Probe for Interactions in the Physiological Denatured State of Chymotrypsin Inhibitor 2
(1997) In Journal of Molecular Biology 269(4). p.611-622- Abstract
- There are four peptidyl-proline bonds in the 64-residue protein chymotrypsin inhibitor 2 (CI2), all of which are in the trans conformation in the native structure. The isomerisation of one or more of these peptidyl-proline bonds to the cis conformation in the denatured state gives rise to heterogeneity, leading to both fast and slow-folding species. The refolding of the fast-folding species, which has all trans peptidyl-proline bonds, is much faster than that of the slow-folding species, which have one or more cis peptidyl-proline bonds. In CI2, the slow-folding species can be classified into two groups by their rates of refolding, temperature-dependence, pH-dependence and [GdmCl]-dependence of the rate constants and the effect of... (More)
- There are four peptidyl-proline bonds in the 64-residue protein chymotrypsin inhibitor 2 (CI2), all of which are in the trans conformation in the native structure. The isomerisation of one or more of these peptidyl-proline bonds to the cis conformation in the denatured state gives rise to heterogeneity, leading to both fast and slow-folding species. The refolding of the fast-folding species, which has all trans peptidyl-proline bonds, is much faster than that of the slow-folding species, which have one or more cis peptidyl-proline bonds. In CI2, the slow-folding species can be classified into two groups by their rates of refolding, temperature-dependence, pH-dependence and [GdmCl]-dependence of the rate constants and the effect of peptidyl-prolyl isomerase on the rate constants. The replacement of Pro6 by Ala removes one of the slow refolding phases, suggesting that the cis peptidyl-Pro6 conformation is solely responsible for one of the slow-folding species. Pro6 is located in a region of the protein where non-random interactions have been found in a series of N-terminal fragments of CI2 (residues 1 to 13, 1 to 25, 1 to 28 and 1 to 40). In addition, NMR studies on a mutant fragment, (1-40)T3A, have confirmed that this non-native interaction is associated with the bulky side-chain of Trp5. The atypical rate of cis to trans isomerisation of the peptidyl-Pro bond is indicative of the presence of a similar hydrophobic cluster in the physiological denatured state of intact CI2. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/126248
- author
- Tan, Yee-Joo ; Oliveberg, Mikael LU ; Otzen, Daniel E and Fersht, Alan R
- organization
- publishing date
- 1997
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- protein, folding, cis-trans
- in
- Journal of Molecular Biology
- volume
- 269
- issue
- 4
- pages
- 611 - 622
- publisher
- Elsevier
- external identifiers
-
- scopus:0031580205
- pmid:9217264
- ISSN
- 1089-8638
- DOI
- 10.1006/jmbi.1997.1043
- language
- English
- LU publication?
- yes
- id
- 0994f48c-943c-49de-ba71-ab1071724815 (old id 126248)
- date added to LUP
- 2016-04-01 16:35:39
- date last changed
- 2025-10-14 11:28:28
@article{0994f48c-943c-49de-ba71-ab1071724815,
abstract = {{There are four peptidyl-proline bonds in the 64-residue protein chymotrypsin inhibitor 2 (CI2), all of which are in the trans conformation in the native structure. The isomerisation of one or more of these peptidyl-proline bonds to the cis conformation in the denatured state gives rise to heterogeneity, leading to both fast and slow-folding species. The refolding of the fast-folding species, which has all trans peptidyl-proline bonds, is much faster than that of the slow-folding species, which have one or more cis peptidyl-proline bonds. In CI2, the slow-folding species can be classified into two groups by their rates of refolding, temperature-dependence, pH-dependence and [GdmCl]-dependence of the rate constants and the effect of peptidyl-prolyl isomerase on the rate constants. The replacement of Pro6 by Ala removes one of the slow refolding phases, suggesting that the cis peptidyl-Pro6 conformation is solely responsible for one of the slow-folding species. Pro6 is located in a region of the protein where non-random interactions have been found in a series of N-terminal fragments of CI2 (residues 1 to 13, 1 to 25, 1 to 28 and 1 to 40). In addition, NMR studies on a mutant fragment, (1-40)T3A, have confirmed that this non-native interaction is associated with the bulky side-chain of Trp5. The atypical rate of cis to trans isomerisation of the peptidyl-Pro bond is indicative of the presence of a similar hydrophobic cluster in the physiological denatured state of intact CI2.}},
author = {{Tan, Yee-Joo and Oliveberg, Mikael and Otzen, Daniel E and Fersht, Alan R}},
issn = {{1089-8638}},
keywords = {{protein; folding; cis-trans}},
language = {{eng}},
number = {{4}},
pages = {{611--622}},
publisher = {{Elsevier}},
series = {{Journal of Molecular Biology}},
title = {{The Rate of Isomerisation of Peptidyl-proline Bonds as a Probe for Interactions in the Physiological Denatured State of Chymotrypsin Inhibitor 2}},
url = {{http://dx.doi.org/10.1006/jmbi.1997.1043}},
doi = {{10.1006/jmbi.1997.1043}},
volume = {{269}},
year = {{1997}},
}