Human platelet vasopressin receptor identification by direct ultraviolet photoaffinity labeling
Tritiated vasopressin ((/sup 3/H)AVP) was directly crosslinked to its human platelet receptor by using an ultraviolet irradiation procedure. After preincubation with (/sup 3/H)AVP, the hydrodynamic parameters of the hormone-receptor complexes solubilized with 3-((3-cholamidopropyl)dimethylammonio)-1-propane sulfonate were derived from Sephacryl S-300 superfine gel filtration and from sucrose density gradient ultracentrifugation experiments. The following values were obtained: Stoke's radius = 5.48 +/- 0.1 nm, apparent sedimentation coefficient = 5.55 +/- 0.1 S, and calculated molecular weight = 132,000. On sodium dodecyl sulfate-8% polyacrylamide slab gel electrophoresis under reducing conditions, (/sup 3/H)AVP preferentially and specifically labeled a 125,000-dalton protein. The labeling of this protein was suppressed by addition of excess cold vasopressin, whereas angiotensin II did not inhibit incorporation of tritiated vasopressin in this protein. These results suggest that direct UV-photoaffinity labelling with (/sup 3/H)AVP is a suitable tool for the purification of the human platelet vasopressin receptor.
- Research Organization:
- Medical College of Ohio, Toledo
- OSTI ID:
- 6025475
- Journal Information:
- J. Biol. Chem.; (United States), Journal Name: J. Biol. Chem.; (United States) Vol. 262:23; ISSN JBCHA
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
AFFINITY
ANGIOTENSIN
ANIMALS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BLOOD PLATELETS
BODY FLUIDS
CARDIOVASCULAR AGENTS
CENTRIFUGATION
CHEMISTRY
DRUGS
ELECTROMAGNETIC RADIATION
ELECTROPHORESIS
FRACTIONATION
GLOBULINS
HORMONES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LABELLING
MAMMALS
MAN
MATERIALS
MEMBRANE PROTEINS
MOLECULAR WEIGHT
ORGANIC COMPOUNDS
PEPTIDE HORMONES
PHOTOCHEMISTRY
PITUITARY HORMONES
PRIMATES
PROTEINS
RADIATIONS
RECEPTORS
SEPARATION PROCESSES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ULTRACENTRIFUGATION
ULTRAVIOLET RADIATION
VASOCONSTRICTORS
VASOPRESSIN
VERTEBRATES