title: Purification and properties of a soluble angiotensin II-binding protein from rabbit liver.
abstract: An angiotensin II-binding activity has been purified almost 3,000-fold to a nearly homogenous state from the 100,000 x g supernatant fraction of rabbit liver. The responsible protein is apparently monomeric since its molecular weight was estimated to be 75,000 in the native state by glycerol gradient centrifugation and in the reduced, denatured state by gel electrophoresis. The Kd and Bmax values of the purified preparation were 7.2 nM and 15.2 nmol of angiotensin II bound per mg of protein, the latter figure agreeing well with the theoretical value of 13.3. Competition experiments with 125I-angiotensin II and unlabeled peptides revealed that the angiotensin antagonist [Sar1,Ala8]angiotensin II (saralasin) and the agonist [des-Asp1]angiotensin II (angiotensin III) were more tightly bound than angiotensin II, whereas angiotensin I and the carboxyl-terminal hexapeptide were less avidly bound. The cardiac peptide, atrial natriuretic factor, also competed for binding to the purified preparation but was about 15-fold less effective than angiotensin II. Although the binding activity was purified in the absence of detergent, a requirement for detergent in the binding reaction emerged during the isolation procedure. Binding by the purified protein exhibited an almost complete dependence upon the presence of detergent, p-chloromercuriphenylsulfonic acid and EDTA.
authors.affiliation: Department of Biochemistry, Cornell University Medical College, New York, New York 10021.
authors.forename: M A
authors.initials: MA
authors.lastname: Kiron
authors.forename: R L
authors.initials: RL
authors.lastname: Soffer
mesh_terms: D000818:Animals; D002845:Chromatography; D003600:Cytosol / Q000032:analysis; D003902:Detergents / Q000494:pharmacology; D007700:Kinetics; D008024:Ligands; D008099:Liver / Q000032:analysis*; D011817:Rabbits; D011945:Receptors, Angiotensin / Q000302:isolation & purification*
chemical_list: D003902:Detergents; D008024:Ligands; D011945:Receptors, Angiotensin