title: Are Caenorhabditis elegans receptors useful targets for drug discovery: pharmacological comparison of tyramine receptors with high identity from C. elegans (TYRA-2) and Brugia malayi (Bm4).
abstract: The biogenic amine, tyramine (TA), modulates a number of key processes in nematodes and a number of TA-specific receptors have been identified. In the present study, we have identified a putative TA receptor (Bm4) in the recently completed Brugia malayi genome and compared its pharmacology to its putative Caenorhabditis elegans orthologue, TYRA-2, under identical expression and assay conditions. TYRA-2 and Bm4 are the most closely related C. elegans and B. malayi BA receptors and differ by only 14aa in the TM regions directly involved in ligand binding. Membranes from HEK-293 cells stably expressing Bm4 exhibited specific, saturable, high affinity, [(3)H]LSD and [(3)H]TA binding with K(d)s of 18.1+/-0.93 and 15.1+/-0.2 nM, respectively. More importantly, both TYRA-2 and Bm4 TA exhibited similar rank orders of potencies for a number of potential tyraminergic ligands. However, some significant differences were noted. For example, chloropromazine exhibited an order of magnitude higher affinity for Bm4 than TYRA-2 (pK(i)s of 7.6+/-0.2 and 6.49+/-0.1, respectively). In contrast, TYRA-2 had significantly higher affinity for phentolamine than Bm4. These results highlight the utility of the nearly completed B. malayi genome and the importance of using receptors from individual parasitic nematodes for drug discovery.
authors.affiliation: Department of Biological Sciences, University of Toledo, 2801 West Bancroft Street, Toledo, OH 43606-3390, USA.
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Smith
authors.forename: Elizabeth B
authors.initials: EB
authors.lastname: Rex
authors.forename: Richard W
authors.initials: RW
authors.lastname: Komuniecki
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017178:Brugia malayi / Q000235:genetics*; D017173:Caenorhabditis elegans / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002746:Chlorpromazine / Q000378:metabolism; D003001:Cloning, Molecular; D006801:Humans; D008024:Ligands; D008238:Lysergic Acid Diethylamide / Q000378:metabolism; D008969:Molecular Sequence Data; D010646:Phentolamine / Q000378:metabolism; D010802:Phylogeny; D017434:Protein Structure, Tertiary / Q000235:genetics; D017982:Receptors, Biogenic Amine / Q000235:genetics* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014439:Tyramine / Q000378:metabolism
chemical_list: D029742:Caenorhabditis elegans Proteins; D008024:Ligands; D017982:Receptors, Biogenic Amine; C502594:TYRA-2 protein, C elegans; C066175:tyramine receptor; D008238:Lysergic Acid Diethylamide; D002746:Chlorpromazine; D014439:Tyramine; D010646:Phentolamine