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Cu(II)-Binding properties of a cytochrome c with a synthetic metal-binding site: His-X_3-His in an α-helix

Todd, Robert J. and Van Dam, Mariana E. and Casimiro, Danilo and Haymore, Barry L. and Arnold, Frances H. (1991) Cu(II)-Binding properties of a cytochrome c with a synthetic metal-binding site: His-X_3-His in an α-helix. Proteins, 10 (2). pp. 156-161. ISSN 0887-3585 . https://resolver.caltech.edu/CaltechAUTHORS:20180822-141010187

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Abstract

A metal‐binding site consisting of two histidines positioned His‐X_3‐His in an α‐helix has been engineered into the surface of Saccharomyces cerevisiae iso‐1‐cytochrome c. The synthetic metal‐binding cytochrome c retains its biological activity in vivo. Its ability to bind chelated Cu(II) has been characterized by partitioning in aqueous two‐phase polymer systems containing a polymer‐metal complex, Cu(II)IDA‐PEG, and by metal‐affinity chromatography. The stability constant for the complex formed between Cu(II)IDA‐PEG and the cytochrome c His‐X3‐His site is 5.3 × 104^ M^(−1), which corresponds to a chelate effect that contributes 1.5 kcal mol^(−1) to the binding energy. Incorporation of the His‐X_3‐His site yields a synthetic metal‐binding protein whose metal affinity is sensitive to environmental conditions that alter helix structure or flexibility.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1002/prot.340100209DOIArticle
ORCID:
AuthorORCID
Arnold, Frances H.0000-0002-4027-364X
Alternate Title:Cu(II)-Binding properties of a cytochrome c with a synthetic metal-binding site: His-X3-His in an α-helix
Additional Information:© 1991 Wiley‐Liss. Manuscript received: 01 November 1990. Manuscript accepted: 18 January 1991.
Subject Keywords:synthetic metalloproteins; protein engineering; iso‐1‐cytochrome c; metal binding
Issue or Number:2
Record Number:CaltechAUTHORS:20180822-141010187
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20180822-141010187
Official Citation:Todd, R. J., Van Dam, M. E., Casimiro, D. , Haymore, B. L. and Arnold, F. H. (1991), Cu(II)‐Binding properties of a cytochrome c with a synthetic metal‐binding site: His‐X3‐His in an α‐helix. Proteins, 10: 156-161. doi:10.1002/prot.340100209
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:89058
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:22 Aug 2018 21:38
Last Modified:03 Oct 2019 20:12

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