Transmembrane helix orientation influences membrane binding of the intracellular juxtamembrane domain in Neu receptor peptides

• Published in: Proceedings of the National Academy of Sciences - PNAS 2013-01, Vol.110 (5), p.1646-1651
• Main Authors: Matsushita, Chihiro, Tamagaki, Hiroko, Miyazawa, Yudai, Aimoto, Saburo, Smith, Steven O., Sato, Takeshi
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino Acid Substitution; Animals; Binding sites; Biochemistry; Biological Sciences; Calmodulin - chemistry; Calmodulin - metabolism; Deuterium; Dimerization; Dimers; Fluorescence; Growth factor receptors; Kinases; Kinetics; Ligands; Lipid Bilayers - chemistry; Lipid Bilayers - metabolism; Lipids; Magnetic Resonance Spectroscopy; Membrane Proteins - chemistry; Membrane Proteins - metabolism; Membranes; Molecular Sequence Data; Mutation; Peptides; Peptides - chemistry; Peptides - genetics; Peptides - metabolism; Protein Binding; Protein Multimerization; Protein Structure, Secondary; Protein Structure, Tertiary; Rats; Receptor, ErbB-2 - chemistry; Receptor, ErbB-2 - genetics; Receptor, ErbB-2 - metabolism; Receptors; Spectrometry, Fluorescence; Spectroscopy
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1215207110

The transmembrane (TM) and juxtamembrane (JM) regions of the ErbB family receptor tyrosine kinases connect the extracellular ligand-binding domain to the intracellular kinase domain. Evidence for the role of these regions in the mechanism of receptor dimerization and activation is provided by TM–JM peptides corresponding to the Neu (or rat ErbB2) receptor. Solid-state NMR and fluorescence spectroscopy show that there are tight interactions of the JM sequence with negatively charged lipids, including phosphatidylinositol 4,5-bisphosphate, in TM–JM peptides corresponding to the wild-type receptor sequence. We observe a release of the JM sequence from the negatively charged membrane surface using peptides containing an activating V664E mutation within the TM domain or in peptides engineered to form TM helix dimers with Val664 in the interface. These results provide the basis of a mechanism for coupling ligand binding to kinase activation in the full-length receptor.