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Central European Journal of Biology

, Volume 6, Issue 2, pp 167–175 | Cite as

Expression of human melanocortin 4 receptor in Saccharomyces cerevisiae

  • Vita IgnatovicaEmail author
  • Ramona Petrovska
  • Davids Fridmanis
  • Janis Klovins
Research Article
  • 48 Downloads

Abstract

The melanocortin 4 receptor (MC4R) is involved in the regulation of energy homeostasis and is known as one of the major hypothalamic regulators of food intake. Several studies have shown that replacement of aspartic acid at position 126 of the MC4R abolishes the ligand binding. We used the modified yeast Saccharomyces cerevisiae strain MMY28 to functionally express the MC4R and characterise the importance of this amino acid for ligand based activation of the receptor. The efficiency of the functional expression system was estimated by activation with αMSH, ACTH and THIQ and compared with cAMP response in mammalian cells. We generated the library of MC4R mutants randomised at the amino acid position 126. Recombinant MC4R clones were screened for the αMSH induced activity in yeast. From 9 different amino acids obtained only the natural aspartic acid displayed the ligand dependent activity of MC4R. The MC4R variants with glutamic acid and leucine at position 126, however, displayed higher background activity than other amino acid substitutions. The results suggest that the yeast expression system is suitable for screening of the MC4R receptor ligands and that the substitution of aspartic acid at position 126 of MC4R by different amino acids functionally inactivates the receptor.

Keywords

G-protein coupled receptors Saccharomyces cerevisiae melanocortin receptors functional screens 

Abbreviations

ACTH

adrenocorticotropin hormone

AT

3-Amino-1,2,4-triazole

GPCR

G protein-coupled receptors

MC4R

melanocortin 4 receptor

MSH

melanocyte-stimulating-hormone

THIQ

tetrahydroisoquinoline

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Copyright information

© © Versita Warsaw and Springer-Verlag Wien 2011

Authors and Affiliations

  • Vita Ignatovica
    • 1
    Email author
  • Ramona Petrovska
    • 1
  • Davids Fridmanis
    • 1
  • Janis Klovins
    • 1
  1. 1.Latvian Biomedical Research and Study CentreRigaLatvia

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