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PMID: 18667691 Published · ppublish English Comparative Study Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S.

Light-activated DNA binding in a designed allosteric protein.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 105 ·No. 31 ·2008-08-05 ·Pages 10709-14

Strickland D, Moffat K, Sosnick TR

Abstract

An understanding of how allostery, the conformational coupling of distant functional sites, arises in highly evolvable systems is of considerable interest in areas ranging from cell biology to protein design and signaling networks. We reasoned that the rigidity and defined geometry of an alpha-helical domain linker would make it effective as a conduit for allosteric signals. To test this idea, we rationally designed 12 fusions between the naturally photoactive LOV2 domain from Avena sativa phototropin 1 and the Escherichia coli trp repressor. When illuminated, one of the fusions selectively binds operator DNA and protects it from nuclease digestion. The ready success of our rational design strategy suggests that the helical "allosteric lever arm" is a general scheme for coupling the function of two proteins.

MeSH Terms
Allosteric Site/genetics Avena/genetics Bacterial Proteins/genetics,metabolism Cryptochromes DNA/genetics,metabolism Flavoproteins/genetics,metabolism Light Models, Molecular Protein Binding Protein Engineering/methods Protein Structure, Secondary/genetics Protein Structure, Tertiary Repressor Proteins/genetics,metabolism
Chemicals
Bacterial Proteins Cryptochromes Flavoproteins Repressor Proteins TRPR protein, E coli DNA
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Strickland Devin
Department of Biochemistry and Molecular Biology and Institute for Biophysical Dynamics, University of Chicago, 929 East 57th Street, Chicago, IL 60637, USA.
Moffat Keith
Sosnick Tobin R
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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
1091-6490
DOI
10.1073/pnas.0709610105
Published
2008-08-05
Epub
2008-00-30
Pages
10709-14
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC2504796
Subset
IM
Grants
NIGMS NIH HHS · R01 GM055694-10 · United States
NIGMS NIH HHS · R01 GM055694-11 · United States
NIGMS NIH HHS · R01 GM055694 · United States
NIGMS NIH HHS · R01 GM055694-12 · United States
NIGMS NIH HHS · R01 GM055694-13 · United States
External Links
PubMed source DOI: 10.1073/pnas.0709610105 PMC Full Text View journal details
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