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Response of Rubredoxin from Pyrococcus furiosus to Environmental Changes: Implications for the Origin of Hyperthermostability

Cavagnero, Silvia and Zhou, Zhi H. and Adams, Michael W. W. and Chan, Sunney I. (1995) Response of Rubredoxin from Pyrococcus furiosus to Environmental Changes: Implications for the Origin of Hyperthermostability. Biochemistry, 34 (31). pp. 9865-9873. ISSN 0006-2960. https://resolver.caltech.edu/CaltechAUTHORS:20171120-141844682

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Abstract

The bases of the hyperthermostability of rubredoxin from Pyrococcus furiosus (RdPf) have been probed by structural perturbations induced by solution pH and ionic strength changes. Comparison of the solution behavior at pH 7 and pH 2, as probed by far- and near-UV circular dichroism, Trp fluorescence emission, l-anilinonaphthalene-8-sulfonate (ANS) binding, and NMR spectroscopy, reveals the presence of only minimal structural variations at room temperature. At pH 2, the protein displays a surprising nearly native-like behavior at high ionic strength while, at low ionic strength, it is capable of strongly binding the hydrophobic probe ANS. All the secondary and tertiary structural features, including the environment of the hydrophobic core, appear to be intact regardless of pH and ionic strength. The apparent "melting" or denaturation temperature at pH 2, however, is 42 °C lower than at pH 7. This is attributed to the perturbation of many electrostatic interactions, including the disruption of all the ion pairs, which is complete at pH 2, as indicated by electrometric pH titrations. The implications of these findings for the origins of the hyperthermostability of rubredoxin are discussed.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1021/bi00031a007DOIArticle
http://pubs.acs.org/doi/abs/10.1021/bi00031a007PublisherArticle
ORCID:
AuthorORCID
Chan, Sunney I.0000-0002-5348-2723
Additional Information:© 1995 American Chemical Society. This work was supported by NIH Grants GM 22432 (S.I.C.) and GM 50736 (M.W.W.A.) from the National Institute of General Medical Sciences, U.S. Public Health Service. Contribution No. 9039 from the Arthur Amos Noyes Laboratories of Chemical Physics. We thank Professor Pamela J. Bjorkman for the use of the program GRASP and Professor Douglas C. Rees for helpful discussion on this study.
Funders:
Funding AgencyGrant Number
NIHGM 22432
NIHGM 50736
National Institute of General Medical SciencesUNSPECIFIED
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Other Numbering System NameOther Numbering System ID
Caltech Arthur Amos Noyes Laboratories of Chemical Physics9039
Issue or Number:31
Record Number:CaltechAUTHORS:20171120-141844682
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20171120-141844682
Official Citation:Response of Rubredoxin from Pyrococcus furiosus to Environmental Changes: Implications for the Origin of Hyperthermostability. Silvia Cavagnero, Zhi H. Zhou, Michael W. W. Adams, and Sunney I. Chan. Biochemistry 1995 34 (31), 9865-9873. DOI: 10.1021/bi00031a007
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83356
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:20 Nov 2017 22:28
Last Modified:03 Oct 2019 19:05

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