Functional differences of invariant and highly conserved residues in the extracellular domain of the glycoprotein hormone receptors.
Multiple interactions exist between human follicle-stimulating hormone (FSH) and the N-terminal hormone-binding fragment of the human FSH receptor (FSHR) extracellular domain (ECD). Binding of the other human glycoprotein hormones to their cognate human receptors (luteinizing hormone receptor (LHR) and thyroid-stimulating hormone receptor (TSHR)) was expected to be similar. This study focuses on amino acid residues in β-strands 2 (Lys(74)), 4 (Tyr(124), Asn(129), and Thr(130)), and 5 (Asp(150) and Asp(153)) of the FSHR ECD identified in the human FSH·FSHR ECD crystal structure as contact sites with the common glycoprotein hormone α-subunit, and on noncontact residues in β-strands 2 (Ser(78)) and 8 (Asp(224) and Ser(226)) as controls. These nine residues are either invariant or highly conserved in LHR and TSHR. Mutagenesis and functional characterization of these residues in all three human receptors allowed an assessment of their contribution to binding and receptor activation. Surprisingly, the six reported α-subunit contact residues of the FSHR ECD could be replaced without significant loss of FSH binding, while cAMP signaling potency was diminished significantly with several replacements. Comparative studies of the homologous residues in LHR and TSHR revealed both similarities and differences. The results for FSH/FSHR were analyzed on the basis of the crystal structure of the FSH·FSHR ECD complex, and comparative modeling was used to generate structures for domains, proteins, and complexes for which no structures were available. Although structural information of hormone-receptor interaction allowed the identification of hormone-receptor contact sites, functional analysis of each contact site was necessary to assess its contribution to hormone binding and receptor activation.
Angelova
Krassimira
K
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia 30602, USA.
de Jonge
Hugo
H
Granneman
Joke C M
JC
Puett
David
D
Bogerd
Jan
J
D002460:Cell Line; D018360:Crystallography, X-Ray; D006801:Humans; D036361:Peptide Hormones / Q000737:chemistry* / Q000378:metabolism; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D018000:Receptors, Peptide / Q000737:chemistry* / Q000378:metabolism; D013329:Structure-Activity Relationship
D036361:Peptide Hormones; D018000:Receptors, Peptide