Purification and partial characterization of canine angiotensinogen.

Abstract

A procedure is described to isolate angiotensinogen (renin substrate) from canine plasma. The isolation procedure resulted in an 800-fold purification with a rate of recovery of approximately 12%. The purified protein has a specific activity of 24 micrograms of angiotensin I/mg protein. The amino terminal amino acid sequence of canine angiotensinogen was found to be identical to that of the horse but to differ from that of human and rat angiotensinogens. Canine angiotensinogen was heterogeneous with respect to molecular weight and isoelectric point. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of pure angiotensinogen revealed two closely spaced bands with apparent molecular weights of 58,000 and 56,000. Chromatofocusing showed four isoforms: Peaks of pure angiotensinogen eluted at pH levels of 4.32, 4.23, 4.15, and 4.04. Isoelectric focusing confirmed the presence of four isoforms. Thus, the purification procedure identified two molecular weight forms and four isoforms of canine angiotensinogen. Isolation of the four isoforms will allow their characterization and the study of their physiological significance.