Glutamate 301 of the mouse gonadotropin-releasing hormone receptor confers specificity for arginine 8 of mammalian gonadotropin-releasing hormone

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dc.contributor.authorFlanagan, C A
dc.contributor.authorBecker, I I
dc.contributor.authorDavidson, J S
dc.contributor.authorWakefield, I K
dc.contributor.authorZhou, W
dc.contributor.authorSealfon, S C
dc.contributor.authorMillar, R P
dc.date.accessioned2021-10-08T07:20:44Z
dc.date.available2021-10-08T07:20:44Z
dc.date.issued1994
dc.description.abstractThe Arg residue at position 8 of mammalian GnRH is necessary for high affinity binding to mammalian GnRH receptors. This requirement has been postulated to derive from an electrostatic interaction of Arg8 with a negatively charged receptor residue. In order to identify such a residue, 8 conserved acidic residues of the mouse GnRH receptor were mutated to isosteric Asn or Gln. Mutant receptors were tested for decreased preference for Arg8-containing ligands by ligand binding and inositol phosphate production. One of the mutants, in which the Glu301 residue was mutated to Gln, exhibited a 56-fold decrease in apparent affinity for mammalian GnRH. The mutant receptor also exhibited decreased affinity for [Lys8]GnRH, but its affinity for [Gln8]GnRH was unchanged compared with the wild type receptor. The apparent affinity of the mutant receptor for the acidic analogue, [Glu8]GnRH, was increased more than 10-fold. The mutant receptor did not, therefore, distinguish mammalian GnRH from analogues with amino acid substitutions at position 8 as effectively as the wild type receptor. This loss of discrimination was specific for the residue at position 8, because the mutant receptor did distinguish mammalian GnRH from analogues with favorable substitutions at positions 5, 6, and 7. These findings show that Glu301 of the GnRH receptor plays a role in receptor recognition of Arg8 in the ligand and are consistent with an electrostatic interaction between these 2 residues.
dc.identifier.apacitationFlanagan, C. A., Becker, I. I., Davidson, J. S., Wakefield, I. K., Zhou, W., Sealfon, S. C., & Millar, R. P. (1994). Glutamate 301 of the mouse gonadotropin-releasing hormone receptor confers specificity for arginine 8 of mammalian gonadotropin-releasing hormone. <i>The Journal of Biological Chemistry</i>, 269(36), 22636 - 22641. http://hdl.handle.net/11427/34993en_ZA
dc.identifier.chicagocitationFlanagan, C A, I I Becker, J S Davidson, I K Wakefield, W Zhou, S C Sealfon, and R P Millar "Glutamate 301 of the mouse gonadotropin-releasing hormone receptor confers specificity for arginine 8 of mammalian gonadotropin-releasing hormone." <i>The Journal of Biological Chemistry</i> 269, 36. (1994): 22636 - 22641. http://hdl.handle.net/11427/34993en_ZA
dc.identifier.citationFlanagan, C.A., Becker, I.I., Davidson, J.S., Wakefield, I.K., Zhou, W., Sealfon, S.C. & Millar, R.P. 1994. Glutamate 301 of the mouse gonadotropin-releasing hormone receptor confers specificity for arginine 8 of mammalian gonadotropin-releasing hormone. <i>The Journal of Biological Chemistry.</i> 269(36):22636 - 22641. http://hdl.handle.net/11427/34993en_ZA
dc.identifier.issn0021-9258
dc.identifier.issn1083-351X
dc.identifier.ris TY - Journal Article AU - Flanagan, C A AU - Becker, I I AU - Davidson, J S AU - Wakefield, I K AU - Zhou, W AU - Sealfon, S C AU - Millar, R P AB - The Arg residue at position 8 of mammalian GnRH is necessary for high affinity binding to mammalian GnRH receptors. This requirement has been postulated to derive from an electrostatic interaction of Arg8 with a negatively charged receptor residue. In order to identify such a residue, 8 conserved acidic residues of the mouse GnRH receptor were mutated to isosteric Asn or Gln. Mutant receptors were tested for decreased preference for Arg8-containing ligands by ligand binding and inositol phosphate production. One of the mutants, in which the Glu301 residue was mutated to Gln, exhibited a 56-fold decrease in apparent affinity for mammalian GnRH. The mutant receptor also exhibited decreased affinity for [Lys8]GnRH, but its affinity for [Gln8]GnRH was unchanged compared with the wild type receptor. The apparent affinity of the mutant receptor for the acidic analogue, [Glu8]GnRH, was increased more than 10-fold. The mutant receptor did not, therefore, distinguish mammalian GnRH from analogues with amino acid substitutions at position 8 as effectively as the wild type receptor. This loss of discrimination was specific for the residue at position 8, because the mutant receptor did distinguish mammalian GnRH from analogues with favorable substitutions at positions 5, 6, and 7. These findings show that Glu301 of the GnRH receptor plays a role in receptor recognition of Arg8 in the ligand and are consistent with an electrostatic interaction between these 2 residues. DA - 1994 DB - OpenUCT DP - University of Cape Town IS - 36 J1 - The Journal of Biological Chemistry LK - https://open.uct.ac.za PY - 1994 SM - 0021-9258 SM - 1083-351X T1 - Glutamate 301 of the mouse gonadotropin-releasing hormone receptor confers specificity for arginine 8 of mammalian gonadotropin-releasing hormone TI - Glutamate 301 of the mouse gonadotropin-releasing hormone receptor confers specificity for arginine 8 of mammalian gonadotropin-releasing hormone UR - http://hdl.handle.net/11427/34993 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/34993
dc.identifier.vancouvercitationFlanagan CA, Becker II, Davidson JS, Wakefield IK, Zhou W, Sealfon SC, et al. Glutamate 301 of the mouse gonadotropin-releasing hormone receptor confers specificity for arginine 8 of mammalian gonadotropin-releasing hormone. The Journal of Biological Chemistry. 1994;269(36):22636 - 22641. http://hdl.handle.net/11427/34993.en_ZA
dc.language.isoeng
dc.publisher.departmentDepartment of Medicine
dc.publisher.facultyFaculty of Health Sciences
dc.sourceThe Journal of Biological Chemistry
dc.source.journalissue36
dc.source.journalvolume269
dc.source.pagination22636 - 22641
dc.source.urihttps://dx.doi.org/10.7196/sajs.718
dc.subject.otherAmino Acid Sequence
dc.subject.otherAnimals
dc.subject.otherArginine
dc.subject.otherCell Line
dc.subject.otherCercopithecus aethiops
dc.subject.otherCloning, Molecular
dc.subject.otherGlutamates
dc.subject.otherGlutamic Acid
dc.subject.otherGonadotropin-Releasing Hormone
dc.subject.otherInositol Phosphates
dc.subject.otherIodine Radioisotopes
dc.subject.otherKinetics
dc.subject.otherMice
dc.subject.otherMolecular Sequence Data
dc.subject.otherMutagenesis, Site-Directed
dc.subject.otherProtein Structure, Secondary
dc.subject.otherRadioligand Assay
dc.titleGlutamate 301 of the mouse gonadotropin-releasing hormone receptor confers specificity for arginine 8 of mammalian gonadotropin-releasing hormone
dc.typeJournal Article
uct.type.publicationResearch
uct.type.resourceJournal Article
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