Deletions of the N‐terminal regions of the human melanocortin receptors

The non‐homologous N‐terminal regions of four human melanocortin (MC) receptors were truncated in order to investigate their putative participation in ligand binding. Eleven constructs were made, where different numbers of residues from the N terminus were deleted. These constructs were used for transient expression experiments in COS cells and analysed by ligand binding. The results show that 27, 25, 28, and 20 amino acids could be deleted from the N terminus of the human MC1, MC3, MC4 and MC5 receptors, respectively, including all potential N‐terminal glycosylation sites in the MC1 and the MC4 receptors, without affecting ligand binding or expression levels. The results indicate that the N‐terminal regions of the human MC1, MC3, MC4 and MC5 receptors, do not play an important role for the ligand binding properties of these receptors.

[1]  J. Wikberg,et al.  Molecular cloning and expression of the human melanocyte stimulating hormone receptor cDNA , 1992, FEBS letters.

[2]  P. Prusis,et al.  A three dimensional model for the interaction of MSH with the melanocortin-1 receptor. , 1995, Biochemical and biophysical research communications.

[3]  C. Strader,et al.  Identification of residues involved in ligand binding to the neurokinin-2 receptor. , 1995, Biochemistry.

[4]  J. Wikberg,et al.  Characterisation of the melanocortin 4 receptor by radioligand binding. , 1996, Pharmacology & toxicology.

[5]  G. Heijne,et al.  Membrane proteins: from sequence to structure. , 1994, Annual review of biophysics and biomolecular structure.

[6]  M. Parmentier,et al.  Molecular cloning of a mouse melanocortin 5 receptor gene widely expressed in peripheral tissues. , 1994, Biochemistry.

[7]  J. Wikberg,et al.  Alternative translation initiation codon for the human melanocortin MC3 receptor does not affect the ligand binding. , 1996, European journal of pharmacology.

[8]  S. Watson,et al.  Molecular cloning, expression, and gene localization of a fourth melanocortin receptor. , 1993, The Journal of biological chemistry.

[9]  T. O'donohue,et al.  The opiomelanotropinergic neuronal and endocrine systems , 1982, Peptides.

[10]  R. Howells,et al.  Studies on mu and delta opioid receptor selectivity utilizing chimeric and site-mutagenized receptors. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[11]  J. Wikberg,et al.  Major pharmacological distinction of the ACTH receptor from other melanocortin receptors. , 1996, Life sciences.

[12]  P. Prusis,et al.  Evidence indicating that the TM4, EL2, and TM5 of the melanocortin 3 receptor Do not participate in ligand binding. , 1996, Biochemical and biophysical research communications.

[13]  V. Chhajlani,et al.  Identification of ligand binding residues in extracellular loops of the melanocortin 1 receptor. , 1996, Biochemical and biophysical research communications.

[14]  D. Segaloff,et al.  Extracellular domain of lutropin/choriogonadotropin receptor expressed in transfected cells binds choriogonadotropin with high affinity. , 1990, The Journal of biological chemistry.

[15]  L. Reichert,et al.  Structure--function relationships of the glycoprotein hormones and their receptors. , 1991, Trends in pharmacological sciences.

[16]  M. Mortrud,et al.  The cloning of a family of genes that encode the melanocortin receptors. , 1992, Science.

[17]  T. Kenakin.  The classification of seven transmembrane receptors in recombinant expression systems. , 1996, Pharmacological reviews.

[18]  L. Miller,et al.  Critical Contributions of Amino-terminal Extracellular Domains in Agonist Binding and Activation of Secretin and Vasoactive Intestinal Polypeptide Receptors. STUDIES OF CHIMERIC RECEPTORS (*) , 1995, The Journal of Biological Chemistry.

[19]  J. Wikberg,et al.  Characterisation of melanocortin receptor subtypes by radioligand binding analysis. , 1995, European journal of pharmacology.

[20]  M. Mortrud,et al.  Localization of the melanocortin-4 receptor (MC4-R) in neuroendocrine and autonomic control circuits in the brain. , 1994, Molecular endocrinology.

[21]  J. Wikberg,et al.  Molecular cloning of a novel human melanocortin receptor. , 1993, Biochemical and biophysical research communications.

[22]  P. Prusis,et al.  Evidence for Alternate Points of Attachment for α-MSH and Its Stereoisomer (Nle4,D-Phe7)-α-MSH at the Melanocortin-1 Receptor , 1994 .

[23]  R. Cone,et al.  Targeted Disruption of the Melanocortin-4 Receptor Results in Obesity in Mice , 1997, Cell.

[24]  J. Wikberg,et al.  Expression of functional melanocortin 1 receptors in insect cells. , 1996, Biochemical and biophysical research communications.

[25]  A. Eberle.  The Melanotropins: Chemistry, Physiology and Mechanisms of Action , 1988 .

[26]  G. von Heijne,et al.  Properties of N-terminal tails in G-protein coupled receptors: a statistical study. , 1995, Protein engineering.

[27]  C. Humblet,et al.  Three-dimensional molecular models of the hMC1R melanocortin receptor: complexes with melanotropin peptide agonists. , 1996, Drug design and discovery.

[28]  V. Chhajlani,et al.  Distribution of cDNA for melanocortin receptor subtypes in human tissues. , 1996, Biochemistry and molecular biology international.

[29]  S. Watson,et al.  Molecular cloning of a novel melanocortin receptor. , 1993, The Journal of biological chemistry.