TY - JOUR
AU - Ewers, Helge
AB - We developed a system for optogenetic release of single molecules in cells. We confined soluble and transmembrane proteins to the Golgi apparatus via a photocleavable protein and released them by short pulses of light. Our method allows for a light dose-dependent delivery of functional proteins to the cytosol and plasma membrane in amounts compatible with single-molecule imaging, greatly simplifying access to single-molecule microscopy of any protein in live cells. We were able to reconstitute ion conductance by delivering BK and LRRC8/volume-regulated anion channels to the plasma membrane. Finally we were able to induce NF-kB signaling in T lymphoblasts stimulated by interleukin-1 by controlled release of a signaling protein that had been knocked out. We observed light-induced formation of functional inflammatory signaling complexes that triggered phosphorylation of the inhibitor of nuclear factor kappa-B kinase only in activated cells. We thus developed an optogenetic method for the reconstitution and investigation of cellular function at the single-molecule level.
TI - An optogenetic method for the controlled release of single molecules
JF - Nature Methods
DO - 10.1038/s41592-024-02204-x
DA - 2024-04-01
UR - https://www.deepdyve.com/lp/springer-journals/an-optogenetic-method-for-the-controlled-release-of-single-molecules-HjvN3zarS9
SP - 666
EP - 672
VL - 21
IS - 4
DP - DeepDyve
ER -