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Rubredoxin Variant Folds without Iron

Strop, Pavel and Mayo, Stephen L. (1999) Rubredoxin Variant Folds without Iron. Journal of the American Chemical Society, 121 (11). pp. 2341-2345. ISSN 0002-7863. http://resolver.caltech.edu/CaltechAUTHORS:20110620-160421544

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Abstract

Pyroccocus furiosus rubredoxin (PFRD), like most studied hyperthermophilic proteins, does not undergo reversible folding. The irreversibility of folding is thought to involve PFRD’s iron-binding site. Here we report a PFRD variant (PFRD-XC4) whose iron binding site was redesigned to eliminate iron binding using a computational design algorithm. PFRD-XC4 folds without iron and exhibits reversible folding with a melting temperature of 82 °C, a thermodynamic stability of 3.2 kcal mol^(-1) at 1 °C, and NMR chemical shifts similar to that of the wild-type protein. This variant should provide a tractable model system for studying the thermodynamic origins of protein hyperthermostability.


Item Type:Article
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http://dx.doi.org/10.1021/ja9834780DOIUNSPECIFIED
http://pubs.acs.org/doi/full/10.1021/ja9834780PublisherUNSPECIFIED
Additional Information:© 1999 American Chemical Society. Received October 1, 1998. Publication Date (Web): March 5, 1999. We thank M. K. Eidness for the wild type Pyroccocus furiosus rubredoxin gene used in this study, S. M. Malakauskas, S. Ross, and C. Sarisky for technical assistance and discussions, and B. I. Dahiyat for discussions. The work was supported by the Howard Hughes Medical Institute (S.L.M.) and a NIH training grant (P.S.).
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Howard Hughes Medical Institute (HHMI)UNSPECIFIED
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Record Number:CaltechAUTHORS:20110620-160421544
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20110620-160421544
Official Citation: Rubredoxin Variant Folds without Iron Pavel Strop and and ,Stephen L. Mayo, Journal of the American Chemical Society 1999 121 (11), 2341-2345
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:24068
Collection:CaltechAUTHORS
Deposited By: Marie Ary
Deposited On:04 Oct 2011 15:49
Last Modified:26 Dec 2012 13:19

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