Isolation of signal transduction complexes using biotin and crosslinking methodologies. Proteomics 2007 Jul;7(14):2371-4
Date
07/12/2007Pubmed ID
17623297DOI
10.1002/pmic.200700219Scopus ID
2-s2.0-34547182806 (requires institutional sign-in at Scopus site) 12 CitationsAbstract
We have developed a strategy to preferentially label the N-terminal alpha-amino groups of intact proteins allowing the internal epsilon-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.
Author List
Freed JK, Smith JR, Li P, Greene ASAuthor
Julie K. Freed MD, PhD Vice Chair, Associate Professor in the Anesthesiology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsBiotin
Cross-Linking Reagents
Protein Binding
Rats
Receptors, Tumor Necrosis Factor
Signal Transduction
Tandem Mass Spectrometry
Tumor Necrosis Factor-alpha