Isolation and characterization of human renin substrate.
Human renin substrate (angiotensinogen) was purified from outdated bank plasma. Purification procedures included ammonium sulfate precipitation, DEAE-cellulose column chromatography, concanavalin A-Sepharose column chromatography, Hydroxylapatite column chromatography preparative isoelectric focusing and Ultrogel AcA 44 gel filtration. The final recovery was 10% and the specific angiotensin I content of 10.5 micrograms/mg of protein was obtained. Polyacrylamide gel and SDS-polyacrylamide gel electrophoresis and analytical ultracentrifugal analyses showed the homogeneity of the purified renin substrate. The molecular weight of 60900 was determined by sedimentation equilibrium studies. Human renin substrate was a glycoprotein containing 13% carbohydrate. Cystine could not be detected on amino acid analysis. The purified renin substrate showed the isoelectric point heterogeneity (pI, 4.6 and 4.9).
D000595:Amino Acid Sequence; D000808:Angiotensinogen / Q000097:blood / Q000302:isolation & purification*; D000809:Angiotensins / Q000302:isolation & purification*; D002241:Carbohydrates / Q000032:analysis; D002845:Chromatography; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D007525:Isoelectric Focusing; D008970:Molecular Weight; D014461:Ultracentrifugation
D000809:Angiotensins; D002241:Carbohydrates; D006023:Glycoproteins; D000808:Angiotensinogen