Large-Scale Purification and Characterization of Human Urinary Kallikrein

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概要

• 論文の詳細を見る

• Kallikrein was highly purified from 1000 liters of fresh pooled urine collected from healthy men by the following procedure : silica gel adsorption, gel filtration on Sephadex G-75,DEAE-Sephadex chromatography, bentonite treatment, affinity chromatography on aprotinin-Sepharose 4B, and rapid gel filtration on a TSK Gel G-3000 SWG column. The final preparation, 17mg of human urinary kallikrein (HUK), was purified 621.6-fold from the crude preparation obtained by silica gel adsorption. One mg of HUK showed activities of 5.58 PNA units toward H-D-Val-Leu-Arg-pNA, and about 1100 kallikrein units in the dog hypotensive assay. HUK thus isolated was found to be pure by means of disc electrophoresis, immunoelectrophoresis, and gel filtration. The following enzymes and bioactive substances were not detected : kininase, urokinase, caseinolytic proteases, blood group substances, substances affecting plasma coagulation, and pyrogens. On isoelectric focusing, HUK separated into microheterogeneous components. The pI's of the dominant components were 3.5,3.8,and 4.1,while the corresponding molecular weights were 5.4×10^4,4.9×10^4,and 4.4×10^4,respectively. An aqueous solution of HUK was stable in the pH range from 6 to 12,and was also stable at 60℃ for 2h at neutral pH. Michaelis constants were from 4.0×10^<-5> to 4.0×10^<-4>M toward five kinds of synthetic substrates conventionally used for HUK assay. Optimal pH's for these substrates and human kininogen were in the alkaline pH range of 7.5 to 11.5.

• 公益社団法人日本薬学会の論文
• 1984-03-25

著者

• 守屋 寛

  Faculty of Pharmaceutical Sciences, Science University of Tokyo

• 長谷川 栄一

  Green Cross Corporation

• 守屋 寛

  Faculty Of Pharmaceutical Science Science University Of Tokyo

• 須山 忠和

  Green Cross Corporation

• 森地 重博

  Green Cross Corporation

• SAKO EIJI

  Green Cross Corporation

関連論文

• Measurement of the Sugar Contents and Their Effect on the Electrophoretical Behaviors of Multiple Forms of Hog Pancreatic Kallikrein
• A Proposal-Use of Combined Assays of Kallikrein Activity Measurement
• Studies on Acidic Arginine Esterase Excreted in Urine. I. Purification and Characterization of Dog Urinary Arginine Esterase
• Purification and Characterization of Dog Urinary Kallikrein
• Action of Various Kallikreins and Related Enzymes on Synthetic Arginine and Lysine Derivatives as Substrates
• Human Renal Kallikrein : Purification and Some Properties
• Studies on Kallikreins.I.Effects of the Gastro-Intestinal System and the Liver on Pancreatic Kallikrein
• Effect of intravenously Applicable Globulin on Phagocytosis of Bacteria by Human Polymorphonuclear Leukocytes
• Synthesis of 4-Methylcoumarin-7-yloxy Tetra-N-acetyl-β-chitotetraoside, a Novel Synthetic Substrate for the Fluorometric Assay of Lysozyme
• Large-Scale Purification and Characterization of Human Urinary Kallikrein
• Studies on Kallikreins.II.Passage of ^I-Labelled Hog Pancreatic Kallikrein across the Rat Intestine

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