In vivo proteomic mapping

摘要

The application discloses methods, materials, and compositions for the labeling of molecules, for example, proteins, in living cells or in subcellular compartments of living cells. In particular, the application relates to proteomic analysis methods; materials and compositions and means based on direct tagging of unknown proteins with tagging enzymes (such as biotin ligase or a peroxidase) within the vicinity of a tagging substrate (such as a tyramide) within living cells, with optional targeting to specific subcellular locations by expression of genetic constructs.

主权项

1. A method for proteomic mapping of a living cell comprising:
contacting a living cell with a tagging enzyme under physiological conditions wherein the tagging enzyme catalyzes a reaction with a tagging substrate resulting in the tagging of proteins within an intracellular spatial location around the enzyme and/or substrate; wherein the tagging enzyme is genetically targeted or is targeted using a targeting sequence such as antibody to a subcellular region; and isolating and analyzing the tagged proteins to create a first proteome map.