Asn(102) of the gonadotropin releasing hormone receptor is a critical determinant of potency for agonists containing C-terminal glycinamide

被引:82
作者
Davidson, JS
McArdle, CA
Davies, P
Elario, R
Flanagan, CA
Millar, RP
机构
[1] UNIV BRISTOL, DEPT MED, BRISTOL BS2 8HW, AVON, ENGLAND
[2] UNIV CAPE TOWN, SCH MED, ENDOCRINE LAB, DEPT MED, CAPE TOWN 7925, SOUTH AFRICA
关键词
D O I
10.1074/jbc.271.26.15510
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We demonstrate a critical role for Asn(102) of the human gonadotropin-releasing hormone (GnRH) receptor in the binding of GnRH, Mutation of Asn(102), located at the top of the second transmembrane helix, to Ala resulted in a 225-fold loss of potency for GnRH. Eight GnRH analogs, all containing glycinamide C termini like GnRH, showed similar losses of potency between 95- and 750-foId for the [Ala(102)]GnRHR, compared with wildtype receptor, In contrast, four GnRH analogs that had ethylamide in place of the C-terminal glycinamide residue, showed much smaller decreases in potency between 2.4- and 11-fold, In comparisons of three agonist pairs, differing only at the C terminus, glycinamide derivatives showed an 11-20-fold greater loss of potency for the mutant receptor than their respective ethylamide derivatives, Thus Asn(102) is a critical determinant of potency specifically for ligands with C terminal glycinamide, while ligands with C-terminal ethylamide are less dependent on Asn(102), These findings indicate a role for Asn(102) in the docking of the glycinamide C terminus and are consistent with hydrogen bonding of the Asn(102) side chain with the C-terminal amide moiety, Taken with previous data, they suggest a region of the GnRH receptor formed by the top of helices 2 and 7 as a binding pocket for the C-terminal part of the ligand.
引用
收藏
页码:15510 / 15514
页数:5
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