Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 56, Issue 46, Pages 14521-14525Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201708151
Keywords
genetic encoding; photocrosslinkers; protein-protein interactions; proteomics
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Funding
- National Natural Science Foundation of China [21521003, 21432002]
- National Key Research and Development Program of China [2016YFA0501500]
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A genetically encoded, multifunctional photocrosslinker was developed for quantitative and comparative proteomics. By bearing a bioorthogonal handle and a releasable linker in addition to its photoaffinity warhead, this probe enables the enrichment of transient and low-abundance prey proteins after intracellular photocrosslinking and prey-bait separation, which can be subject to stable isotope dimethyl labeling and mass spectrometry analysis. This quantitative strategy (termed isoCAPP) allowed a comparative proteomic approach to be adopted to identify the proteolytic substrates of an E. coli protease-chaperone dual machinery DegP. Two newly identified substrates were subsequently confirmed by proteolysis experiments.
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