期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 53, 期 18, 页码 4717-4720出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201310969
关键词
chemical inducers of dimerization; fusion proteins; photocleavable linkers; photolysis; protein-protein interactions
资金
- Swiss National Science Foundation [205320-138302, 205320-143699]
- ESF EuroMEMBRANE Programme [31EM30-126143]
- Novartis (Jubilee) Foundation
- Swiss National Science Foundation (SNF) [205320_143699, 205320_138302, 31EM30-126143] Funding Source: Swiss National Science Foundation (SNF)
Chemical inducers of dimerization (CIDs) have been developed to orchestrate protein dimerization and translocation. Here we present a novel photocleavable HaloTag- and SNAP-tag-reactive CID (MeNV-HaXS) with excellent selectivity and intracellular reactivity. Excitation at 360nm cleaves the methyl-6-nitroveratryl core of MeNV-HaXS. MeNV-HaXS covalently links HaloTag- and SNAP-tag fusion proteins, and enables targeting of selected membranes and intracellular organelles. MeNV-HaXS-mediated translocation has been validated for plasma membrane, late endosomes, lysosomes, Golgi, mitochondria, and the actin cytoskeleton. Photocleavage of MeNV-HaXS liberates target proteins and provides access to optical manipulation of protein relocation with high spatiotemporal and subcellular precision. MeNV-HaXS supports kinetic studies of protein dynamics and the manipulation of subcellular enzyme activities, which is exemplified for Golgi-targeted cargo and the assessment of nuclear import kinetics.
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