Human platelet vasopressin receptor identification by direct ultraviolet photoaffinity labeling

Journal Article · Sat Aug 15 00:00:00 EDT 1987 · J. Biol. Chem.; (United States)

OSTI ID:6025475

Thibonnier, M

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), Vol. 262:23
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
RECEPTORS
FRACTIONATION
LABELLING
VASOPRESSIN
AFFINITY
ANGIOTENSIN
BLOOD PLATELETS
ELECTROPHORESIS
MAN
MOLECULAR WEIGHT
PHOTOCHEMISTRY
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ULTRACENTRIFUGATION
ULTRAVIOLET RADIATION
ANIMALS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARDIOVASCULAR AGENTS
CENTRIFUGATION
CHEMISTRY
DRUGS
ELECTROMAGNETIC RADIATION
GLOBULINS
HORMONES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MATERIALS
MEMBRANE PROTEINS
ORGANIC COMPOUNDS
PEPTIDE HORMONES
PITUITARY HORMONES
PRIMATES
PROTEINS
RADIATIONS
SEPARATION PROCESSES
VASOCONSTRICTORS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques