title: Studies on the role of amino acids 38-45 in the expression of a functional thyrotropin receptor.
abstract: We previously reported that deletion or substitution of a unique eight-amino acid tract (residues 38-45) in the extracellular domain of the human TSH receptor led to the loss of specific ligand binding to the surface of transfected cells. In the present study we analyzed this region in more detail. Using site-directed mutagenesis of the TSH receptor cDNA, we substituted amino acid residues 38-45, either in three overlapping groups of four amino acids each or individually. The resultant TSH receptor mutant cDNAs were stably transfected into Chinese hamster ovary cells, and the cells were tested for their TSH-binding ability. Our data demonstrate that amino acid residues 38-40 and 42-45 in this region of the human TSH receptor can be substituted without alteration in receptor function and are, therefore, not critical in forming or maintaining the TSH-binding site. However, substitution of Cys41, either alone or together with adjacent amino acids, leads to the loss of TSH binding to its receptor. These data suggest a central role for the amino acid in position 41 in preserving the biological function of the TSH receptor.
authors.affiliation: Thyroid Molecular Biology Unit, Veterans Administration Medical Center, San Francisco, California 94121.
authors.forename: H L
authors.initials: HL
authors.lastname: Wadsworth
authors.forename: D
authors.initials: D
authors.lastname: Russo
authors.forename: Y
authors.initials: Y
authors.lastname: Nagayama
authors.forename: G D
authors.initials: GD
authors.lastname: Chazenbalk
authors.forename: B
authors.initials: B
authors.lastname: Rapoport
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D006224:Cricetinae; D015870:Gene Expression / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D011989:Receptors, Thyrotropin / Q000737:chemistry / Q000235:genetics*; D013329:Structure-Activity Relationship; D014162:Transfection / Q000235:genetics
chemical_list: D011989:Receptors, Thyrotropin