1 research outputs found
Optochemical Control of Protein Localization and Activity within Cell-like Compartments
We report inducible dimerization
strategies for controlling protein
positioning, enzymatic activity, and organelle assembly inside synthetic
cell-like compartments upon photostimulation. Using a photocaged TMP-Haloligand
compound, we demonstrate small molecule and light-induced dimerization
of DHFR and Haloenzyme to localize proteins to a compartment boundary
and reconstitute tripartite sfGFP assembly. Using photocaged rapamycin
and fragments of split TEV protease fused to FRB and FKBP, we establish
optical triggering of protease activity inside cell-size compartments.
We apply light-inducible protease activation to initiate assembly
of membraneless organelles, demonstrating the applicability of these
tools for characterizing cell biological processes in vitro. This
modular toolkit, which affords spatial and temporal control of protein
function in a minimal cell-like system, represents a critical step
toward the reconstitution of a tunable synthetic cell, built from
the bottom up