title: Analysis of epidermal growth factor receptor dimerization by BS³ cross-linking.
abstract: Dimerization of receptor tyrosine kinases is a well-characterized process. It is imperative for the activation of many receptors, including the epidermal growth factor receptor (EGFR). EGFR has been shown to be regulated by a number of factors, including lipid raft localization. For example, alteration of the lipid raft localization of EGFR has been demonstrated to modify receptor dimerization. This protocol describes an assay to quantify EGFR dimers using BS(3) cross-linking. BS(3) cross-linking is well suited for this purpose because of its length, water solubility, and membrane impermeability. Although this protocol is written specifically for EGFR, the assay can be extrapolated in order to characterize dimerization of other receptor tyrosine kinases.
authors.affiliation: Program in Integrative Nutrition and Complex Diseases and the Center Translational Environmental Health Research, Texas A&M University, College Station, TX, 77843, USA.
authors.forename: Harmony F
authors.initials: HF
authors.lastname: Turk
authors.forename: Robert S
authors.initials: RS
authors.lastname: Chapkin
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002461:Cell Line, Transformed; D003106:Colon / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D021962:Membrane Microdomains / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D055503:Protein Multimerization; D013388:Succinimides / Q000737:chemistry*
chemical_list: D003432:Cross-Linking Reagents; D013388:Succinimides; D004815:Epidermal Growth Factor; C035760:bis(sulfosuccinimidyl)suberate; C541485:EGFR protein, mouse; D066246:ErbB Receptors