Massachusetts Institute of Technology

Jeffrey Daniel Martell

Alice Y Ting

Peng Zou

Hyun-Woo Rhee

An imaging method utilizing a split peroxidase is described herein. Imaging methods involve contacting a cell with a split peroxidase and a substrate thereof to allow conversion of a substrate into a product via an enzymatic reaction catalyzed by the reconstitute split peroxidase. Also disclosed herein are split peroxidases, related products and kits.

Split peroxidases and methods of use

An imaging method comprising expressing in cells a Class I heme peroxidase, which optionally is fused with a protein of interest or a cellular localization signal peptide, and contacting the cells with a substrate of the Class I heme peroxidase to allow conversion of the substrate into a product via an oxidation reaction catalyzed by the Class I heme peroxidase, wherein the product releases a signal detectable by a microscope such as an electron microscope. Also disclosed herein are monomeric mutants of a Class I heme peroxidase and mutants of the enzyme that exhibit elevated enzymatic activity as compared to the corresponding wild-type counterpart.

Cytosolically-active peroxidases as reporters for microscopy

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.

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Somerville, MA, United States of America

Jeffrey Daniel Martell