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Isolation of signal transduction complexes using biotin and crosslinking methodologies
Author(s) -
Freed Julie K.,
Smith Julia R.,
Li Peigang,
Greene Andrew S.
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200700219
Subject(s) - biotinylation , lysis , biotin , signal transduction , transduction (biophysics) , chemistry , reagent , biochemistry , receptor , membrane , cell surface receptor , combinatorial chemistry , organic chemistry
We have developed a strategy to preferentially label the N‐terminal α‐amino groups of intact proteins allowing the internal ϵ‐amino groups to remain free to react with chemical crosslinking reagents. The convergence of these methodologies allows biotinylated ligands to bind to their receptors within the cell membrane followed by removal of the crosslinked complex from cell lysate. This technique allows for the isolation of protein complexes in an MS‐compatible system, thus providing a tool for furthering our understanding of signal transduction.