Smith Katherine A, Rex Elizabeth B, Komuniecki Richard W
Department of Biological Sciences, University of Toledo, 2801 West Bancroft Street, Toledo, OH 43606-3390, USA.
Mol Biochem Parasitol. 2007 Jul;154(1):52-61. doi: 10.1016/j.molbiopara.2007.04.004. Epub 2007 Apr 13.
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.
生物胺酪胺(TA)可调节线虫体内的一些关键过程,并且已经鉴定出多种TA特异性受体。在本研究中,我们在最近完成的马来布鲁线虫基因组中鉴定出一种假定的TA受体(Bm4),并在相同的表达和检测条件下,将其药理学特性与其假定的秀丽隐杆线虫直系同源物TYRA-2进行了比较。TYRA-2和Bm4是秀丽隐杆线虫和马来布鲁线虫中关系最密切的生物胺受体,在直接参与配体结合的跨膜区域仅相差14个氨基酸。稳定表达Bm4的HEK-293细胞膜表现出特异性、可饱和、高亲和力的[(3)H]麦角酸二乙酰胺(LSD)和[(3)H]TA结合,解离常数(K(d))分别为18.1±0.93和15.1±0.2 nM。更重要的是,TYRA-2和Bm4对多种潜在的酪胺能配体都表现出相似的效价顺序。然而,也注意到了一些显著差异。例如,氯丙嗪对Bm4的亲和力比对TYRA-2高一个数量级(pK(i)分别为7.6±0.2和6.49±0.1)。相反,TYRA-2对酚妥拉明的亲和力比Bm4高得多。这些结果突出了近乎完成的马来布鲁线虫基因组的实用性,以及使用来自单个寄生线虫的受体进行药物发现的重要性。