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PMID: 22388287 Published · epublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

TULIPs: tunable, light-controlled interacting protein tags for cell biology.

Nature methods ·Vol. 9 ·No. 4 ·2012-03-04 ·Pages 379-84

Strickland D, Lin Y, Wagner E, Hope CM, Zayner J, Antoniou C, Sosnick TR, Weiss EL, Glotzer M

Abstract

Naturally photoswitchable proteins offer a means of directly manipulating the formation of protein complexes that drive a diversity of cellular processes. We developed tunable light-inducible dimerization tags (TULIPs) based on a synthetic interaction between the LOV2 domain of Avena sativa phototropin 1 (AsLOV2) and an engineered PDZ domain (ePDZ). TULIPs can recruit proteins to diverse structures in living yeast and mammalian cells, either globally or with precise spatial control using a steerable laser. The equilibrium binding and kinetic parameters of the interaction are tunable by mutation, making TULIPs readily adaptable to signaling pathways with varying sensitivities and response times. We demonstrate the utility of TULIPs by conferring light sensitivity to functionally distinct components of the yeast mating pathway and by directing the site of cell polarization.

MeSH Terms
Animals Avena/chemistry Cell Biology Cell Polarity Enzyme Activation Kinetics Lasers Light Mitogen-Activated Protein Kinases/metabolism Models, Molecular Mutation PDZ Domains Phototropins/chemistry,genetics,metabolism Protein Binding/genetics,radiation effects Protein Engineering Protein Transport/radiation effects Proteins/chemistry,genetics,metabolism Saccharomyces cerevisiae/cytology,enzymology
Chemicals
Phototropins Proteins Mitogen-Activated Protein Kinases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Strickland Devin
Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, Illinois, USA.
Lin Yuan
Wagner Elizabeth
Hope C Matthew
Zayner Josiah
Antoniou Chloe
Sosnick Tobin R
Weiss Eric L
Glotzer Michael
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Article Info
Journal
Nature methods
Abbr.
Nat Methods
ISSN
1548-7105
DOI
10.1038/nmeth.1904
Published
2012-03-04
Epub
2012-00-04
Pages
379-84
Language
English
Region
United States
NLM ID
101215604
PMCID
PMC3444151
Subset
IM
Grants
NIGMS NIH HHS · R01 GM088668 · United States
NIGMS NIH HHS · GM088668 · United States
External Links
PubMed source DOI: 10.1038/nmeth.1904 PMC Full Text View journal details
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