PMID: 78Dec 5, 1975

Studies on rat renal cortical cell kallikrein. I. Separation and measurement

Biochimica Et Biophysica Acta
T Kaizu, H S Margolius

Abstract

A technique has been developed to separate and measure kallikrein in a heterogeneous population of rat renal cortical cells in suspension. After rat kidneys were perfused in situ in anaesthetized rats, viable, counted cortical cell suspensions were obtained. Cells were suspended in a sucrose/Tris buffer containing 0.5% deoxycholate, homogenized, centrifuged, dialyzed, and gel filtered on Sephadex G-25. Column chromatography on DEAE-cellulose resulted in a single peak of esterase activity between 0.20 to 0.25 M NaCl/sodium phosphate buffer. Subsequent elution yielded an alkaline esterase which was identical to kallikrein isolated from rat urine, insofar as pH optimum, effects of inhibitors, bioassay activity and immunological properties were concerned. Calculated yields were about 70% of the total esterase activity present in the parent cell homogenates. Recoveries of a purified rat urinary kallikrein added to the cell homogenates, the DEAE-cellulose columns, or the eluates from the columns ranged from 83-108% (mean 96%). Using this technique, it was found that the amount of kallikrein activity present in non-incubated renal cortical cells ranged from 0.6-10(-2) to 4.6 - 10(-2) alpha-N-tosyl-L-arginine methyl ester (Tos-Arg-OMe)...Continue Reading

Citations

Jan 1, 1981·Pflügers Archiv : European journal of physiology·K TomitaF Sakai
Jan 1, 1978·General Pharmacology·K NustadJ V Pierce
Jun 1, 1980·The Journal of Clinical Investigation·H S Margolius, J Chao
Nov 1, 2011·Clinical and Experimental Nephrology·Lawrence G PalmerGustavo Frindt
Jan 30, 1978·Archives of Biochemistry and Biophysics·J Chao
Dec 15, 1982·Biochemical and Biophysical Research Communications·M TakaokaS Morimoto
Jul 1, 1979·Biochemical Pharmacology·J Chao, H S Margolius
Dec 29, 2000·Japanese Journal of Pharmacology·T FujitaM Majima
Mar 1, 1987·Kidney International·M WeinbergR J Solomon
Nov 1, 1981·Clinics in Endocrinology and Metabolism·K Abe
Apr 1, 1979·Circulation Research·N G Levinsky
Sep 1, 1981·Acta Pharmacologica Et Toxicologica·U B Olsen, E Eilertsen
Mar 17, 2021·AACN Advanced Critical Care·Maureen Scarboro, Karen A McQuillan
Sep 1, 1992·The American Journal of Physiology·S AvigdorJ Bouhnik

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