title: Synthesis and biological activity of a biotinylated vasopressin analog.
abstract: To study vasopressin receptor-mediated endocytosis using electronmicroscopy methods and to develop avidin affinity columns for receptor purification, we synthesized and tested the biological properties of a biotinylated vasopressin (VP) analog [1-(2-mercapto) propionic acid] 8-[lysine-N6-biotin] VP (B-MLVP). B-MLVP was prepared by coupling biotin to the epsilon amine of the lysine residue in [1-(2-mercapto) propionic acid] 8-(lysine) VP (MLVP). The structure of HPLC purified B-MLVP was confirmed by fast atom bombardment mass spectrometry. B-MLVP effectively competed for arginine vasopressin (AVP) binding sites in canine renal plasma membranes on the surface of LLC-PK1 kidney cells. Dissociation constants of 15 nM and 202 nM were calculated from the results of competition binding assays conducted with membranes and cells, respectively. B-MLVP stimulated adenylate cyclase activity and elevated cellular 3',5',cyclic-AMP (cAMP) content in a manner similar to AVP, indicating it is an agonist of VP action in renal tissue. These observations indicate that B-MLVP is an agonist of VP action and may be used to study renal VP receptors by employing avidin coupled to various reporter groups.
authors.affiliation: Department of Medicine, Mayo Clinic and Foundation, Rochester, MN 55905.
authors.forename: W H
authors.initials: WH
authors.lastname: Lutz
authors.forename: J M
authors.initials: JM
authors.lastname: Londowski
authors.forename: R
authors.initials: R
authors.lastname: Kumar
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive / Q000502:physiology; D001710:Biotin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D004285:Dogs; D066298:In Vitro Techniques; D007679:Kidney Medulla / Q000378:metabolism; D008236:Lypressin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011945:Receptors, Angiotensin / Q000378:metabolism; D017483:Receptors, Vasopressin
chemical_list: D011945:Receptors, Angiotensin; D017483:Receptors, Vasopressin; C066225:vasopressin, ((2-mercapto)propionic acid)(1)-(Lys-N(6)-biotin)(8)-; D008236:Lypressin; D001710:Biotin; D000242:Cyclic AMP; D000262:Adenylyl Cyclases