TY - JOUR
T1 - Conserved mammalian gonadotropin-releasing hormone receptor carboxyl terminal amino acids regulate ligand binding, effector coupling and internalization
AU - Brothers, Shaun P.
AU - Janovick, Jo Ann
AU - Maya-Nunez, Guadalupe
AU - Cornea, Anda
AU - Han, Xin Bing
AU - Conn, P. Michael
N1 - Funding Information:
We thank Diane Ryles for helping with the manuscript. This study was supported by NIH grants HD-19899, RR-00163, HD-18185, G.M.N. received support from Fogarty Grant TW/HD00668.
PY - 2002/4/25
Y1 - 2002/4/25
N2 - The mammalian gonadotropin-releasing hormone receptor (GnRHR), with 327 amino acids, is among the smallest G protein coupled receptors identified. Absent from this receptor is the cytoplasmic tail, characteristic of other members of this superfamily, which frequently mediates desensitization and down-regulation. The fifteen carboxyl terminal residues in the mammalian GnRHR are absolutely conserved, suggesting important roles for these residues. In the current study, mutations of the mammalian GnRHR were made to study the carboxyl terminus. The receptor mutant GnRHR(Ser326Ala) was reduced in ligand affinity (117% reduction compared to wild type (wt)), while receptor numbers and internalization remained unchanged. GnRHR(Ser326Tyr) was decreased in effector coupling, while ligand affinity remained unchanged compared to wt. These studies also show that, while mutation of Ser326 caused a change in ligand binding and effector coupling, truncation at this residue (GnRHR[des326-327]) had no measurable effect on GnRHR ligand binding, effector coupling or internalization, functions which appear to require different structural determinants than expression and routing. Removal of all three carboxyl terminal residues (Phe325, Ser326 and Leu327) or mutation of the receptor (GnRHR[Phe325Ala]) caused a complete loss of measurable ligand binding and effector coupling, clearly suggesting an unexplained role for Phe325.
AB - The mammalian gonadotropin-releasing hormone receptor (GnRHR), with 327 amino acids, is among the smallest G protein coupled receptors identified. Absent from this receptor is the cytoplasmic tail, characteristic of other members of this superfamily, which frequently mediates desensitization and down-regulation. The fifteen carboxyl terminal residues in the mammalian GnRHR are absolutely conserved, suggesting important roles for these residues. In the current study, mutations of the mammalian GnRHR were made to study the carboxyl terminus. The receptor mutant GnRHR(Ser326Ala) was reduced in ligand affinity (117% reduction compared to wild type (wt)), while receptor numbers and internalization remained unchanged. GnRHR(Ser326Tyr) was decreased in effector coupling, while ligand affinity remained unchanged compared to wt. These studies also show that, while mutation of Ser326 caused a change in ligand binding and effector coupling, truncation at this residue (GnRHR[des326-327]) had no measurable effect on GnRHR ligand binding, effector coupling or internalization, functions which appear to require different structural determinants than expression and routing. Removal of all three carboxyl terminal residues (Phe325, Ser326 and Leu327) or mutation of the receptor (GnRHR[Phe325Ala]) caused a complete loss of measurable ligand binding and effector coupling, clearly suggesting an unexplained role for Phe325.
KW - G protein coupling
KW - GnRH receptor
KW - Gonadotropin-releasing hormone
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U2 - 10.1016/S0303-7207(02)00040-0
DO - 10.1016/S0303-7207(02)00040-0
M3 - Article
C2 - 11997175
AN - SCOPUS:0037171443
SN - 0303-7207
VL - 190
SP - 19
EP - 27
JO - Molecular and Cellular Endocrinology
JF - Molecular and Cellular Endocrinology
IS - 1-2
ER -