Melanocortin 4 receptors interact with antimicrobial frog peptide analogues.
We have developed fluorescence polarization (FP) assays of human melanocortin 4 receptor (MC4R) in 384-well microtiter plates using TAMRA-NDP-MSH as a tracer. The rank order of potency of agonists and antagonists agrees well relative to the published assays: SHU9119>MTII>NDP alphaMSH>alphaMSH. We have screened libraries of Korean plant extracts and frog peptide analogues in search of MC4R ligands using FP assays and cell-based CRE luciferase reporter assays. We report that FLGFLFKVASK, FLGWLFKVASK, FLGALFKWASK, and FLGWLFKWASK are the peptide analogues, which bind to human MC4R receptor with good affinity in vitro. FLGWLFKVASK and FLGWLFKWASK stimulated CRE-driven reporter gene via MC4R. In luciferase reporter assays, they possess the pharmacological and functional profiles of full agonists. We demonstrate the interaction of MC4R with 11-residue antimicrobial peptides derived from the Korean frog, Rana rugosa. The results suggest that MC4R interacts promiscuously with bioactive analogues of antimicrobial peptide, gaegurin-5.
School of Agricultural Biotechnology, Seoul National University, San 56-1 Sillim-dong, Gwanak-gu, Seoul 151-742, Republic of Korea.
Ernest U
EU
Do
Eun Bae
EB
Jo
Gyu
G
Choi
Long Zhu
LZ
Piao
Jaekyoon
J
Shin
Min-Duk
MD
Seo
Su-Jin
SJ
Kang
Bong-Jin
BJ
Lee
Kang Ho
KH
Kim
Jae Bum
JB
Kim
Su-il
SI
Kim
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000494:pharmacology*; D001667:Binding, Competitive; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D005454:Fluorescence Polarization; D017930:Genes, Reporter; D006801:Humans; D008024:Ligands; D009074:Melanocyte-Stimulating Hormones / Q000494:pharmacology*; D009842:Oligopeptides / Q000494:pharmacology*; D010936:Plant Extracts / Q000494:pharmacology; D011898:Ranidae; D044105:Receptor, Melanocortin, Type 4 / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011994:Recombinant Proteins / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism; D000521:alpha-MSH / Q000031:analogs & derivatives* / Q000494:pharmacology
D023181:Antimicrobial Cationic Peptides; D008024:Ligands; C470072:MC4R protein, human; D009842:Oligopeptides; D010936:Plant Extracts; D044105:Receptor, Melanocortin, Type 4; D011994:Recombinant Proteins; C105974:acetyl-norleucyl(4)-(aspartyl(5)-histidyl(6)-phenylalanyl(7)-arginyl(8)-tryptophyl(9)-lysyl(10))cyclo-alpha-MSH(4-10)amide; C103272:SHU 9119; D000521:alpha-MSH; C027756:MSH, 4-Nle-7-Phe-alpha-; D009074:Melanocyte-Stimulating Hormones; D000242:Cyclic AMP