L-phenylalanine and NPS R-467 synergistically potentiate the function of the extracellular calcium-sensing receptor through distinct sites.
The extracellular calcium (Ca(2+)(o))-sensing receptor (CaR) can be potentiated by allosteric activators including calcimimetics and l-amino acids. In this study, we found that many mutations had differential effects on the functional modulation of the CaR by these two allosteric activators, supporting the idea that these modulators act through distinct sites. 10 mm l-phenylalanine and 1 microm NPS R-467, submaximal doses of the two agents, each elicited similar modulation of R185Q. However, there are different relative potencies for these two modulators with some receptors being more responsive to l-phenylalanine and others being more responsive to NPS R-467. The responsiveness of the CaR to Ca(2+)(o) appears to be essential to observe the potentiating action of l-phenylalanine but not of NPS R-467 on the receptor. NPS R-467 reduces the Hill coefficients of the wild-type as well as mutant receptors, suggesting that engagement of all Ca(2+) binding sites is not required when the receptor is activated by NPS R-467. In contrast, l-phenylalanine has little effect on the Hill coefficients of mutant receptors. The two-site model is further supported by the observation that these two classes of modulators exert a synergistic effect on CaRs with inactivating mutations that are responsive to both modulators.
Department of Medicine, Endocrine-Hypertension Division, Brigham and Women's Hospital and Harvard Medical School, 221 Longwood Avenue, Boston, MA 02115, USA.
Zaixiang
Z
Zhang
Yongfeng
Y
Jiang
Stephen J
SJ
Quinn
Karen
K
Krapcho
Edward F
EF
Nemeth
Mei
M
Bai
D000494:Allosteric Regulation; D000814:Aniline Compounds / Q000494:pharmacology*; D001665:Binding Sites; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D004357:Drug Synergism; D006801:Humans; D010649:Phenylalanine / Q000494:pharmacology*; D044169:Receptors, Calcium-Sensing; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000187:drug effects* / Q000502:physiology
D000814:Aniline Compounds; C111654:NPS R-467; D044169:Receptors, Calcium-Sensing; D011956:Receptors, Cell Surface; D010649:Phenylalanine; D002118:Calcium