Open AccessLOVTRAP: A Versatile Method to Control Protein Function with Light
Author(s) -
Wang Hui,
Hahn Klaus M.
Publication year - 2016
Publication title -
current protocols in cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.149
H-Index - 38
eISSN - 1934-2616
pISSN - 1934-2500
DOI - 10.1002/cpcb.12
Subject(s) - biophysics , membrane , chemistry , membrane protein , irradiation , function (biology) , nanotechnology , microbiology and biotechnology , biochemistry , materials science , biology , physics , nuclear physics
We describe a detailed procedure for the use of LOVTRAP, an approach to reversibly sequester and release proteins from cellular membranes using light. In the application described here, proteins that act at the plasma membrane are held at mitochondria in the dark, and reversibly released by irradiation. The technique relies on binding of an engineered Zdk domain to a LOV2 domain, with affinity <30 nM in the dark and >500 nM upon irradiation between 400 and 500 nm. LOVTRAP can be applied to diverse proteins, as it requires attaching only one member of the Zdk/LOV2 pair to the target protein, and the other to the membrane where the target protein is to be sequestered. Light‐induced protein release occurs in less than a second, and the half‐life of return can be adjusted using LOV point mutations (∼2 to 500 sec). © 2016 by John Wiley & Sons, Inc.