Mechanism of dimerization of the human melanocortin 1 receptor
Section snippets
Materials and methods
Materials. Igepal CA-630, BSA, EDTA, PMSF, iodoacetamide and bicinchoninic acid were from Sigma (St. Louis, MO). The synthetic αMSH analogue [Nle4, D-Phe7] αMSH (NDP-MSH) was from Calbiochem (Darmstadt, Germany). The radioligand [125I]NDP-MSH, specific activity 2000 Ci/mmol, and a cAMP radioimmunoassay kit were from Amersham Pharmacia Biotech (Little Chalfont, Buckinghamshire, UK). Other reagents were from Merck (Darmstadt, Germany) or Prolabo (Barcelona, Spain).
Expression constructs and
Results and discussion
Dimerization of the MC1R was previously demonstrated by co-immunoprecipitation studies [17]. The mechanisms of GPCR dimerization are often incompletely understood, but may involve: (i) coiled-coil interactions between the C-terminal domains, (ii) covalent intersubunit disulfide bonding, and (iii) the swapping of large receptor domains to yield functional units that comprise structural elements from each partner. We have analyzed the contribution of these three types of intersubunit interactions
Acknowledgments
Supported by Grant SAF2006-11206 from the Comisión Interministerial de Ciencia y Tecnología (CICYT), Spain, and FEDER funds (European Community). BLS-L and ABP-O hold fellowships from the Ministry of Education (Spain), and CH from the Fundación Séneca (CARM). PTZ was a UNESCO-L’OREAL fellow for Women in Science in 2005 and a fellow from the Puglia Region (Italy). We thank Prof. R. Peñafiel and A. López-Contreras (Department of Biochemistry and Molecular Biology, University of Murcia) for gift
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