Unique helical conformation of the fourth cytoplasmic loop of the CB1 cannabinoid receptor in a negatively charged environment

Christy R.R. Grace, Sudha M. Cowsik, Joong Youn Shim, William J. Welsh, Allyn C. Howlett

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

The proximal portion of the C-terminus of the CB1 cannabinoid receptor is a primary determinant for G-protein activation. A 17 residue proximal C-terminal peptide (rodent CB1 401-417), the intracellular loop 4 (IL4) peptide, mimicked the receptor's G-protein activation domain. Because of the importance of the cationic amino acids to G-protein activation, the three-dimensional structure of the IL4 peptide in a negatively charged sodium dodecyl sulfate (SDS) micellar environment has been studied by two-dimensional proton nuclear magnetic resonance (2D 1H NMR) spectroscopy and distance geometry calculations. Unambiguous proton NMR assignments were carried out with the aid of correlation spectroscopy (DQF-COSY and TOCSY) and nuclear Overhauser effect spectroscopy (NOESY and ROESY) experiments. The distance constraints were used in torsion angle dynamics algorithm for NMR applications (DYANA) to generate a family of structures which were refined using restrained energy minimization and dynamics. In water, the IL4 peptide prefers an extended conformation, whereas in SDS micelles, 310-helical conformation is induced. The predominance of 310-helical domain structure in SDS represents a unique difference compared with structure in alternative environments, which can significantly impact global electrostatic surface potential on the cytoplasmic surface of the CB1 receptor and might influence the signal to the G-proteins.

Original languageEnglish (US)
Pages (from-to)359-368
Number of pages10
JournalJournal of Structural Biology
Volume159
Issue number3
DOIs
StatePublished - Sep 2007

All Science Journal Classification (ASJC) codes

  • Structural Biology

Keywords

  • Cell surface 7-transmembrane receptors
  • G-protein coupled receptors
  • Signal transduction mechanisms
  • Sodium dodecyl sulfate (SDS) micelles
  • Two-dimensional proton nuclear magnetic resonance (2D H NMR) spectroscopy

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