Purification of prorennin and production of its antibody.
Prorennin, i.e. the zymogen of rennin, was extracted from fresh calf stomach and purified by ammonium sulfate precipitation, ion exchange chromatography and gel filtration. The purified prorennin was homogeneous as judged by SDS-polyacrylamide gel electrophoresis. A specific antibody to the purified prorennin was induced in rabbits. In a double diffusion test and on immunoelectrophoresis the antibody gave a single precipitin line when run against crude and pure zymogen preparations. In the double diffusion test, the antibody also reacted with a purified rennin but not with gastric proteases (pepsin and pepsinogen) or other proteases (trypsin and Mucor rennin). When the antibody was tested against prorennin and rennin in the double diffusion test, a spur was formed at the junction of the precipitin lines against prorennin and rennin. This result confirms that the two proteins share an antigenically common structure in their molecules and that the activation segment of prorennin contains at least one antigenic determinant.
D000818:Animals; D000918:Antibody Specificity; D002417:Cattle; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D012085:Chymosin / Q000276:immunology* / Q000302:isolation & purification; D004792:Enzyme Precursors / Q000276:immunology* / Q000302:isolation & purification; D000939:Epitopes / Q000276:immunology; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D008297:Male; D011817:Rabbits / Q000276:immunology
D004792:Enzyme Precursors; D000939:Epitopes; C032443:prorennin; D012085:Chymosin