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Photo-crosslinking: An Emerging Chemical Tool for Investigating Molecular Networks in Live Cells |
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| Authors: | Pratyush Kumar Mishra Chang-Mo Yoo Dr Eunmi Hong Prof?Dr Hyun Woo Rhee |
| Affiliation: | 1. Department of Chemistry, Seoul National University, Gwanak-ro 1, Gwanak-gu, Seoul, 08826 Republic of Korea
Department of Chemistry, Ulsan National Institute of Science and Technology (UNIST), 50 UNIST-gil, Eonyang-eup, Ulju-gun, Ulsan, 44919 Republic of Korea;2. Department of Chemistry, Seoul National University, Gwanak-ro 1, Gwanak-gu, Seoul, 08826 Republic of Korea;3. New Drug Development Center, Daegu-Gyeongbuk Medical Innovation Foundation (DGMIF), 360-4 Dongnae-dong, Dong-gu, Daegu, 41061 Republic of Korea |
| Abstract: | Studying protein–protein interactions (PPIs) is useful for understanding cellular functions and mechanisms. Evaluating these PPIs under conditions as similar as possible to native conditions can be achieved using photo-crosslinking methods because of their on-demand ability to generate reactive species in situ by irradiation with UV light. Various fusion tag, metabolic incorporation, and amber codon suppression approaches using various crosslinkers containing aryl azide, benzophenone, and diazirines have been applied in live cells. Mass spectrometry and immunological techniques are used to identify crosslinked proteins based on their capture transient and context-dependent interactions. Herein we discuss various incorporation methods and crosslinkers that have been used for interactome mapping in live cells. |
| Keywords: | enzymatic labeling photo-crosslinking protein–protein interactions proximity labeling unnatural amino acids |