The catecholamine binding site of the beta-adrenergic receptor is formed by juxtaposed membrane-spanning domains.
The catecholamine binding domain of the turkey erythrocyte beta-adrenergic receptor was mapped by determining the sites of covalent labeling of the purified receptor by two beta-adrenergic photoaffinity reagents, [125I]iodocyanopindolol-diazirine (ICYP-da) and [125I] iodoazidobenzylpindolol (IABP). Both labels were incorporated at two separate sites. By sequencing a labeled peptide, one site of labeling was found to lie at Trp330 in the extracellular half of the seventh membrane span. This position is homologous to the retinal attachment site in rhodopsin. The second labeled site was isolated on an 8000-Da peptide and immunoprecipitated using sequence-directed antibodies. This site lies in membrane spans 3-5. Labeling of the two sites was equal using ICYP-da and 3-10-fold greater in the span 7 site using IABP. These data indicate that the catecholamine binding site is formed from the juxtaposition of span 7 and spans 3-5 in a tertiary structure probably similar to that of rhodopsin.
Wong
S K
SK
Department of Pharmacology, Southwestern Graduate School of Biomedical Sciences, University of Texas Health Science Center, Dallas 75235-9041.
Slaughter
C
C
Ruoho
A E
AE
Ross
E M
EM
D000345:Affinity Labels / Q000378:metabolism; D000818:Animals; D001386:Azides / Q000378:metabolism; D001665:Binding Sites; D002395:Catecholamines / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D003978:Diazomethane / Q000378:metabolism; D020120:Iodocyanopindolol; D010449:Peptide Mapping; D010777:Photochemistry; D010869:Pindolol / Q000031:analogs & derivatives / Q000378:metabolism; D011943:Receptors, Adrenergic, beta / Q000378:metabolism*; D012243:Rhodopsin / Q000378:metabolism; D014422:Turkeys
D000345:Affinity Labels; D001386:Azides; D002395:Catecholamines; D011943:Receptors, Adrenergic, beta; D003978:Diazomethane; C029987:1-(indol-4-yloxy)-3-(1-(4-azido-3-iodophenyl)-2-isobutylamine)-2-propanol; D020120:Iodocyanopindolol; D012243:Rhodopsin; D010869:Pindolol