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PMID: 8346210 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Genetic fusion of subunits of a dimeric protein substantially enhances its stability and rate of folding.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 90 ·No. 15 ·1993-08-01 ·Pages 7010-4

Liang H, Sandberg WS, Terwilliger TC

Abstract

The gene V protein of bacteriophage f1 is a single-stranded DNA and RNA-binding protein composed of two identical subunits. We have constructed single-chain variants of the protein using short peptide linkers of five or six amino acids to connect the carboxyl terminus of one monomer to the amino terminus of the second monomer. The resulting subunit-fusion gene V proteins were found to bind single-stranded DNA nearly as tightly as the wild-type protein. Denaturation measurements show that the subunit-fusion gene V proteins are 5 kcal/mol (1 kcal = 4.18 kJ) more stable than the wild-type protein at a protein concentration of 10 microM. The rate of unfolding of the protein is essentially unaffected by the fusion of monomeric subunits, whereas the rate of folding is greatly enhanced. Our results suggest a simple way of obtaining a substantial thermodynamic stabilization for some oligomeric proteins.

MeSH Terms
Coliphages/chemistry DNA-Binding Proteins/chemistry Macromolecular Substances Protein Denaturation Protein Structure, Secondary Recombinant Fusion Proteins/chemistry Viral Proteins/chemistry
Chemicals
DNA-Binding Proteins Macromolecular Substances Recombinant Fusion Proteins Viral Proteins gene V protein, Enterobacteria phage f1
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Liang H
Department of Biochemistry and Molecular Biology, University of Chicago, IL 60637.
Sandberg W S
Terwilliger T C
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34 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.90.15.7010
Published
1993-08-01
Pages
7010-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC47065
Subset
IM
Grants
NIGMS NIH HHS · GM 38714 · United States
External Links
PubMed source DOI: 10.1073/pnas.90.15.7010 PMC Full Text View journal details
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