Purification and characterization of renin binding protein (RnBP) from porcine kidney

Journal of Biochemistry
S TakahashiY Miyake

Abstract

Renin binding protein (RnBP) was purified from porcine kidney using pepstatin affinity column chromatography, DEAE-Sepharose column chromatography, gel filtration on Ultrogel-AcA 34, aminohexyl-Sepharose 4B column chromatography, and high performance liquid chromatography (HPLC) on TSK-gel G-3000 SW. The purified preparation was homogeneous as judged by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, polyacrylamide disc gel electrophoresis and isoelectric focusing on polyacrylamide gel. The isoelectric point was at pH 4.85, and the apparent molecular weight of RnBP was estimated to be 42,000 by SDS-polyacrylamide gel electrophoresis. The preparation did not show any renin activity and was stable for 30 min at 37 degrees C between pH 5.0 and 9.0 or on storage for 4 weeks at 4 degrees C or -80 degrees C. The activity of renin was greatly inhibited by RnBP. From the kinetic analysis of the inhibition we roughly estimated the dissociation constant between renin and RnBP to be about 0.2 nM, assuming that the stoichiometry in the complex, i.e., high molecular weight (HMW) renin, is one to one, and that the complex is inactive. The inhibitory activity of RnBP was lost by acidification at pH 3.0 and the activity of ren...Continue Reading

References

Dec 9, 2008·Bioscience, Biotechnology, and Biochemistry·Saori TakahashiSeihan Yamada
Apr 5, 2011·Journal of the Renin-angiotensin-aldosterone System : JRAAS·Rogério FerreiraPaulo Bayard Gonçalves
Sep 1, 1991·Hypertension·T Inagami
Oct 7, 2006·Journal of Biochemistry·Saori TakahashiToshihiro Sugiyama

Related Concepts

Polyacrylamide Gels
Complex (molecular entity)
Immune Sera
Precipitins
Acidification - ActCode
Pepstatins
Kidney
Structure of Cortex of Kidney
Binding Protein
Gel Chromatography

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