Identification and sequence of a binding site peptide of the beta 2-adrenergic receptor.

Published

Journal Article

p-(Bromoacetamido)benzyl-1-[125I]iodocarazolol (125I-pBABC) is a potent derivative of the beta-adrenergic receptor antagonist p-aminobenzylcarazolol. Treatment of the receptor with 125I-pBABC results in efficient covalent incorporation of the ligand into the receptor binding site. Extensive degradation of 125I-pBABC-labeled beta 2-adrenergic receptor with either cyanogen bromide or Staphylococcus aureus V8 protease results in specifically labeled fragments having Mr's of about 1600 and 3500, respectively. Because the primary structure of the beta 2-adrenergic receptor is known, and these proteolytic reagents are highly sequence specific, the site of 125I-pBABC incorporation may be deduced from the sizes of the specifically labeled fragments. Thus the fragment generated by cyanogen bromide cleavage corresponds to residues 83-96, a region of 14 amino acids included in the second membrane spanning domain (helix II) of the beta 2-adrenergic receptor. This assignment was confirmed by direct amino acid sequencing of this labeled fragment, though the actual amino acid modified could not be determined. These data permit the assignment of a part of the hormone binding region of the beta 2-adrenergic receptor.

Full Text

Duke Authors

Cited Authors

  • Dohlman, HG; Caron, MG; Strader, CD; Amlaiky, N; Lefkowitz, RJ

Published Date

  • March 22, 1988

Published In

Volume / Issue

  • 27 / 6

Start / End Page

  • 1813 - 1817

PubMed ID

  • 2837273

Pubmed Central ID

  • 2837273

International Standard Serial Number (ISSN)

  • 0006-2960

Digital Object Identifier (DOI)

  • 10.1021/bi00406a002

Language

  • eng

Conference Location

  • United States