title: Oligomers of ERBB3 have two distinct interfaces that differ in their sensitivity to disruption by heregulin.
abstract: ErbB receptors associate in a ligand-dependent or -independent manner, and overexpression of epidermal growth factor receptor (ErbB1) or ErbB2 results in ligand-independent activation. Ligand-independent activation is poorly understood, and dimerization alone is not sufficient for activation. ErbB receptors also form higher order oligomers, but the mechanism of oligomer formation and their contribution to signaling are not known. The kinase-deficient ErbB3 as well as its extracellular domains are particularly prone to ligand-independent oligomerization, and oligomers are destabilized by binding of the ligand heregulin. In contrast, ligand binding facilitates heterodimerization with ErbB2 and is expected to stabilize an extended conformation of the ErbB3 extracellular domain (ECD) in which the dimerization interface is exposed. In the absence of ligand, ErbB3 can adopt a closed conformation that is held together by an intramolecular tether. We used a constitutively extended form of the ErbB3-ECD to analyze the conformation of the ECD in oligomers and the mechanism of oligomer disruption by heregulin. The extended conformation of the ECD forms oligomers more readily, suggesting the crystallographically defined dimer interface is one of the interfaces involved in oligomerization. Heregulin destabilizes oligomeric complexes but not dimers, which are neither stabilized nor disrupted by ligand binding, indicating a distinct second interface in oligomers of ErbB3. Cross-linking and activation studies on membrane-embedded ErbB3/ErbB2 chimeras confirm this dual effect of heregulin. Most of the ErbB3-ECD on the cell surface is apparently kept in an open conformation through oligomerization, and the resulting oligomers adopt a conformation representing a state of reduced activity.
authors.affiliation: Department of Medicine, Hematology-Oncology, UCLA, Los Angeles, California 90095, USA.
authors.forename: Kian
authors.initials: K
authors.lastname: Kani
authors.forename: Carmen M
authors.initials: CM
authors.lastname: Warren
authors.forename: Catherine S
authors.initials: CS
authors.lastname: Kaddis
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Loo
authors.forename: Ralf
authors.initials: R
authors.lastname: Landgraf
mesh_terms: D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003432:Cross-Linking Reagents / Q000494:pharmacology; D018360:Crystallography, X-Ray; D018076:DNA, Complementary / Q000378:metabolism; D019281:Dimerization; D006801:Humans; D047468:Immunoprecipitation; D008024:Ligands; D008958:Models, Molecular; D020890:Neuregulin-1 / Q000737:chemistry*; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D020893:Receptor, ErbB-3 / Q000737:chemistry* / Q000378:metabolism; D012685:Sepharose / Q000737:chemistry; D015398:Signal Transduction; D021241:Spectrometry, Mass, Electrospray Ionization; D013997:Time Factors; D014443:Tyrosine / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D018076:DNA, Complementary; D008024:Ligands; D020890:Neuregulin-1; D014443:Tyrosine; D012685:Sepharose; D020893:Receptor, ErbB-3