title: Analysis of the Role of the C-Terminal Tail in the Regulation of the Epidermal Growth Factor Receptor.
abstract: The ∼230-residue C-terminal tail of the epidermal growth factor receptor (EGFR) is phosphorylated upon activation. We examined whether this phosphorylation is affected by deletions within the tail and whether the two tails in the asymmetric active EGFR dimer are phosphorylated differently. We monitored autophosphorylation in cells using flow cytometry and found that the first ∼80 residues of the tail are inhibitory, as demonstrated previously. The entire ∼80-residue span is important for autoinhibition and needs to be released from both kinases that form the dimer. These results are interpreted in terms of crystal structures of the inactive kinase domain, including two new ones presented here. Deletions in the remaining portion of the tail do not affect autophosphorylation, except for a six-residue segment spanning Tyr 1086 that is critical for activation loop phosphorylation. Phosphorylation of the two tails in the dimer is asymmetric, with the activator tail being phosphorylated somewhat more strongly. Unexpectedly, we found that reconstitution of the transmembrane and cytoplasmic domains of EGFR in vesicles leads to a peculiar phenomenon in which kinase domains appear to be trapped between stacks of lipid bilayers. This artifactual trapping of kinases between membranes enhances an intrinsic functional asymmetry in the two tails in a dimer.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, California, USA Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Erika
authors.initials: E
authors.lastname: Kovacs
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, California, USA Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Rahul
authors.initials: R
authors.lastname: Das
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, California, USA Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Medicine, Washington University School of Medicine, St. Louis, Missouri, USA.
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Collier
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, California, USA Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Aaron
authors.initials: A
authors.lastname: Cantor
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, California, USA Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Yongjian
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, California, USA Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Kathryn
authors.initials: K
authors.lastname: Wong
authors.affiliation: Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Amar
authors.initials: A
authors.lastname: Mirza
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, California, USA Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Tiago
authors.initials: T
authors.lastname: Barros
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Patricia
authors.initials: P
authors.lastname: Grob
authors.affiliation: Cardiovascular Research Institute and Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, California, USA.
authors.forename: Natalia
authors.initials: N
authors.lastname: Jura
authors.affiliation: Department of Medicine, Washington University School of Medicine, St. Louis, Missouri, USA.
authors.forename: Ron
authors.initials: R
authors.lastname: Bose
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, California, USA Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, California, USA Department of Chemistry, University of California, Berkeley, Berkeley, California, USA Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA kuriyan@berkeley.edu.
authors.forename: John
authors.initials: J
authors.lastname: Kuriyan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D018360:Crystallography, X-Ray; D004789:Enzyme Activation / Q000235:genetics; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure*; D005434:Flow Cytometry; D057809:HEK293 Cells; D006801:Humans; D056004:Molecular Dynamics Simulation; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion / Q000235:genetics
chemical_list: D066246:ErbB Receptors