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Science. 2015 Apr 17;348(6232):344-7. doi: 10.1126/science.aaa0445.

Protein structure. Engineering of a superhelicase through conformational control.

Author information

1
Physics Department and Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
2
Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
3
Physics Department and Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA. Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA. Howard Hughes Medical Institute, University of Illinois, Urbana, IL 61801, USA. tjha@illinois.edu.

Abstract

Conformational control of biomolecular activities can reveal functional insights and enable the engineering of novel activities. Here we show that conformational control through intramolecular cross-linking of a helicase monomer with undetectable unwinding activity converts it into a superhelicase that can unwind thousands of base pairs processively, even against a large opposing force. A natural partner that enhances the helicase activity is shown to achieve its stimulating role also by selectively stabilizing the active conformation. Our work provides insight into the regulation of nucleic acid unwinding activity and introduces a monomeric superhelicase without nuclease activities, which may be useful for biotechnological applications.

PMID:
25883358
PMCID:
PMC4417355
DOI:
10.1126/science.aaa0445
[Indexed for MEDLINE]
Free PMC Article

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