Affinity capture of (Arg sup 8 )vasopressin-receptor complex using immobilized antisense peptide

Journal Article · Wed May 01 00:00:00 EDT 1991 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

OSTI ID:5934180

Lu, Feng Xian; Aiyar, N; Chaiken, I ^[1]

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1. SmithKline Beecham, King of Prussia, PA (United States)

Solubilized noncovalent complexes of (Arg{sup 8})-vasopressin (AVP) with receptor proteins from rat liver membranes were isolated by selective binding to silica-immobilized antisense (AS) peptide. The affinity chromatographic support was prepared with a chemically synthesized AS peptide whose sequence is encoded by the AS DNA corresponding to the 20 amino-terminal residues of the AVP bovine neurophysin II biosynthetic precursor (pro-AVP/BNPII-(20-1)), region that includes the AVP sequence at residues 1-9. The AS peptide-AVP interaction mechanism hypothesized, contact by hydropathic complementarity at multiple sites along the peptide chains, led to the prediction that AVP bound to its receptor would still have enough free surface to interact with immobilized AS peptide. To test this prediction of a three-way interaction, ({sup 3}H)AVP-receptor was obtained as a solubilized, partially purified fraction from rat liver membrane. Covalently crosslinked ({sup 3}H)AVP complex also was bound to the AS peptide column; binding was blocked by competition with unlabeled AVP in the elution buffer. Since the AVP-linked 31- and 38-kDa proteins have the same apparent molecular mass on SDS/PAGE as found previously by photoaffinity labeling, the authors conclude that the AS peptide column has affinity-captured AVP-receptor complexes. The 15-kDa protein appears to be an active AVP-receptor fragment of one or both of the larger proteins. It is generally concluded that immobilized AS peptides may be useful to isolate peptide and protein receptor complexes in other systems as well.

• OSTI ID: 5934180
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Vol. 88:9; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
RECEPTORS
CROSS-LINKING
VASOPRESSIN
ARGININE
IMMOBILIZED ENZYMES
LIVER
PEPTIDES
AMINO ACIDS
BODY
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
DIGESTIVE SYSTEM
GLANDS
HORMONES
MEMBRANE PROTEINS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PEPTIDE HORMONES
PITUITARY HORMONES
POLYMERIZATION
PROTEINS
550201* - Biochemistry- Tracer Techniques