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Enrichment of Specifically Labeled Proteins by an Immobilized Host Molecule
Author(s) -
Murray James,
Sim Jaehwan,
Oh Kyunghoon,
Sung Gihyun,
Lee Ara,
Shrinidhi Annadka,
Thirunarayanan Ayyavu,
Shetty Dinesh,
Kim Kimoon
Publication year - 2017
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201611894
Subject(s) - streptavidin , chemistry , biotin , proteomics , click chemistry , chemical biology , conjugated system , molecule , adamantane , biochemistry , combinatorial chemistry , organic chemistry , gene , polymer
Chemical proteomics relies primarily on click‐chemistry‐based protein labeling and biotin‐streptavidin enrichment, but these techniques have inherent limitations. Enrichment of intracellular proteins using a totally synthetic host–guest complex is described, overcoming the problem associated with the classical approach. We achieve this by affinity‐based protein labeling with a target‐specific probe molecule conjugated to a high‐affinity guest (suberanilohydroxamic acid–ammonium‐adamantane; SAHA‐Ad) and then enriching the labeled species using a cucurbit[7]uril bead. This method shows high specificity for labeled molecules in a MDA‐MB‐231 breast cancer cell lysate. Moreover, this method shows promise for labeling proteins in live cells.