CHARACTERIZATION OF A SYNTHETIC PEPTIDE ANTIBODY RECOGNIZING RAT-KIDNEY RENIN AND PRORENIN

A fourteen amino acid peptide from the cDNA predicted rat renin amino acid sequence was synthesized and used to produce a polyclonal antibod y, The ability of this antibody to recognize rat renin was characteriz ed and compared to that of a previously tested polyclonal anti-rat ren in antibody raised against purified rat renin to verify its usefulness as a reagent in renin processing. The anti-renin peptide antibody and anti-rat renin antibody were evaluated for specificity of recognition of rat renin by immunohistochemical staining and immunoblotting. The ability of the anti-renin peptide antibody to inhibit renin enzymatic activity and to immunoprecipitate rat recombinant protein was also tes ted. Anti-renin peptide antibody and anti-rat renin antibody had ident ical recognition of rat renin and prorenin in immunohistochemical stai ns of rat kidney section and immunoblots. However the anti-renin pepti de antibody does not inhibit renin enzymatic activity nor does it immu noprecipitate recombinant rat prorenin. We have produced and character ized an antirenin peptide antibody which recognizes denaturated rat re nin and prorenin in a way that is identical to that of an antibody rai sed against native rat renin. The anti-renin peptide antibody does not recognize nondenatured renin because it did not inhibit renin enzymat ic activity nor was it able to immunoprecipitate renin or prorenin. Th e technique of using synthetic peptides to produce antibody recognizin g at renin is useful because low yields of native renin purified from rat kidney are generally insufficient for antibody production, This li mits the widespread availability of antibodies that recognize rat reni n. Alternative ways of producing antibodies which recognize rat renin and prorenin will help to facilitate renin angiotensin research.