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K Kizuki

Publications and source records attributed to K Kizuki.

At least 37 records · Page 2Linked to original sources
PubMed1989

Characterization of N-linked oligosaccharides of human urinary kallikrein molecules.

The micro-heterogeneity due to varied N-linked oligosaccharides of both active- and pro-types of human urinary kallikrein (HUK) in normal subjects and some patients were investigated by the methods of serial lectin affinity chromatography and crossed affino-immunoelectrophoresis. In the case of both types of normal HUK, the species carrying tri- and/or tetra-antennary oligosaccharide(s), corefucosylated bi-antennary oligosaccharide(s), and bi-antennary oligosaccharides containing outer galactose residues and an N-acetylglucosamine residue linked beta 1,4 to a beta-linked mannose residue (bisecting N-acetylglucosamine residue) amounted to approximately 36, 33 and 17% of the total of each type of HUK, respectively. On the other hand, in some diseases, i.e. essential hypertension, Bartter's syndrome and acute pancreatitis, alterations of the chromatographic and electrophoretic patters were observed and are assumed to correspond to glycosylation changes in each HUK molecule.

Chromatography, Affinity↗
PubMed1989

Characterization of human pancreatic kallikrein.

Human pancreatic kallikrein (H.Panc.K.) was purified from human pancreas by ion exchange chromatography on DEAE-cellulose, affinity chromatographies on p-aminobenzamidine Sepharose 6B and aprotinin aminocellulofine, followed by gel filtration on Sephacryl S-200. The final preparation had a specific activity of 9.2 AU/A280 (AU; amidase unit for H-Pro-Phe-Arg-MCA) and its N-terminal sequence coincided with the reported sequence for H.Panc.K.. In HPLC (gel filtration), one symmetrical peak corresponding to a molecular weight of 48,000 was obtained. In SDS-PAGE without 2-mercaptoethanol (2-ME), one band corresponding to a molecular weight of 52,000 was obtained, but with 2-ME, 2 bands, 52,000 and 30,000, were obtained. Km value for MCA was 4.9 x 10(-2) mM. Proteinase inhibitor specificities of H.Panc.K. were the same as those of human urinary kallikrein (HUK) and hog pancreatic kallikrein (hog Panc.K.), while anti-HUK antibody inhibited the activities of H.Panc.K. and HUK, but not that of hog Panc.K.. From the analysis of affinity for concanavalin A (Con A) and erythroagglutinating phytohemagglutinin (E-PHA), the carbohydrate parts of H.Panc.K. are relatively rich in biantennary complex type sugar chains with bisecting GlcNAc compared with those of human salivary kallikrein (H.Saliv.K.) and HUK.

Amino Acid Sequence↗
PubMed1989

Enzyme-linked immunosorbent assays for human tissue kallikrein and analysis of immunoreactive kallikrein in the plasma by them.

In order to study immunoreactive tissue kallikrein (TK) in human plasma, three kinds of enzyme-linked immunosorbent assay (ELISA) systems for human urinary kallikrein (HUK) were developed. All three types of TK in the plasma, i.e., active kallikrein, prokallikrein and kallikrein-alpha 1 antitrypsin (alpha 1 AT) complex, could be measured by the competitive ELISA (C-ELISA). However, the sandwich ELISA (S-ELISA) showed more strict specificity. Namely, both active and prokallikreins were able to be measured by the S-ELISA, while kallikrein-alpha 1 AT complex was hardly measurable. Measurement of this kallikrein-alpha 1 AT complex was made possible by using horseradish peroxidase (HRP)-labeled anti-alpha 1 AT IgG instead of HRP-labeled anti-HUK Fab' as the second antibody (HS-ELISA). Using these ELISAs we studied TK in plasma and the following observations were made. 1) The greater part of TK in the plasma was found in a form of a complex with alpha 1 AT. 2) The amount of free type TK was very small and the most part of this type TK in normal plasma was prokallikrein.

Chromatography, Affinity↗
PubMed1989

Distribution of angiotensin I converting enzyme in male reproductive systems of various vertebrates and properties of the genital enzymes.

The distribution of the angiotensin I converting enzyme (EC 3.4.15.1, ACE) in male reproductive systems of various vertebrates including non-mammalian species and properties of the genital ACEs were studied. In such mammals as rat, dog and pig, it has been found that ACE activity is only distributed in testis and epididymis (especially in the epididymal semen), but not in accessory sex glands such as prostate, seminal vesicle and coagulating gland. In the rat and dog, most of or all epididymal ACE has been found to resemble testicular ACEs rather than pulmonary ACE in molecular weight. Besides the studies on mammals, it has been found that the enzymes having characteristics of ACE are present in the genital systems of such lower vertebrates as bird (domestic fowl) and fish (carp).

Animals↗
PubMed1989

[Purification of human pancreatic kallikrein and organ-specificities of human glandular kallikreins].

Human pancreatic kallikrein (H. Panc. K.) was purified from human pancreas by serial liquid chromatographies. The final preparation had a specific activity of 9.2 AU/A280 (AU: amidase unit for H-Pro-Phe-Arg-MCA) and its N-terminal sequence coincided with the reported sequence determined from cloned cDNA analysis. In HPLC (gel filtration), one symmetrical peak corresponding to a molecular weight of 48,000 was obtained. In SDS-PAGE without 2-mercaptoethanol, one band corresponding to a molecular weight of 52,000 was obtained. Protease inhibitor specificities of H. Panc. K. were the same as those of human urinary kallikrein (HUK) and hog pancreatic kallikrein (hog Panc. K.), while anti-HUK rabbit antibody inhibited the activities of H. Panc. K. and HUK, but not that of hog Panc. K. From the analysis of affinity for concanavalin A and erythroagglutinating phytohemagglutinin, the carbohydrate parts of H. Panc. K. are relatively rich in bi-(or multi-) antennary complex type sugar chains with bisecting GlcNAc compared with those of human salivary kallikrein and HUK. These findings will be a help to clarify the physiological and pathophysiological roles of H. Panc. K. in the pancreas and pancreatic diseases, especially in acute pancreatitis.

Carbohydrate Conformation↗
PubMed1986

An inactive form of kallikrein in human urine.

An inactive form of human urinary kallikrein (inactive HUK) was highly purified from fresh urine collected from healthy men. Inactive HUK was separated from the active kallikrein (HUK) initially presents in the urine by affinity chromatography on a column of aprotinin immobilized on Sepharose 4B and further purified by gel filtration, ion-exchange chromatography and immunoaffinity chromatography on an anti-HUK antibody immobilized Sepharose 4B column. Inactive HUK was rapidly activated by a trace amount of trypsin. While, plasmin, urokinase, thrombin and chymotrypsin caused no activation of inactive HUK. The molecular weights of inactive HUK and HUK were estimated to be 4.8 X 10(4) and 4.5 X 10(4), respectively. The molecular weight of active HUK generated from inactive HUK by the action of trypsin (HUK'') was almost the same as that of HUK. The mobility of inactive HUK was slightly slower than that of HUK on both immunoelectrophoresis and polyacrylamide gel disc electrophoresis. On the other hand, the electrophoretic mobility of HUKK'' was almost the same as that of HUK. These two types of active HUK had no significant difference in the Km values for H-Pro-Phe-Arg-MCA hydrolysis and inhibition profiles by various protease inhibitors and anti-HUK antibody. Inactive HUK was unable to be measured by the direct radioimmunoassay (RIA) but HUK" generated by the action of trypsin could be measured by the RIA.

Humans↗
PubMed1986

Studies on carbohydrate structure and immunological properties of human urinary kallikreins.

Some studies on carbohydrates structure and immunological properties of human urinary kallikreins purified from the human urine of healthy men (HUK) were investigated. A general technique for fractionating asparagine-linked oligosaccharides of HUK was applied. This involves serial chromatographies on concanavalin A-Sepharose 4B and other lectins bound Agarose. Asparagine-linked main oligosaccharides of both active and inactive types of HUK were mixture of tri- or tetra-antennary complex oligosaccharide having GlcNAC beta 1-4Man beta residue (bisecting GlcNAc) (type I-2), bi-antennary complex oligosaccharide having core Fuc alpha 1-6GlcNAc residue (type III-3), tri- or tetra-antennary complex oligosaccharides having Gal beta 1-4GlcNAc beta 1-4 (Gal beta 1-4GlcNAc beta 1-2)Man alpha-residue (type I-1-A-a) etc. Besides these structural studies, the method to probe the heterogeneous profiles of HUK due to the different structures of carbohydrate chains bound, was devised without the complete purification of urinary kallikrein even with individual subject of not only active but also inactive forms of the kallikrein.

Animals↗
PubMed1986

Rat testicular angiotensin I converting enzyme: purification and comparison with rat pulmonary enzyme.

Enzymological properties of rat testicular angiotensin I converting enzyme (RT-ACE) were compared with those of rat pulmonary angiotensin I converting enzyme (RP-ACE). The molecule of RT-ACE was different from that of RP-ACE with respect to the molecular weight, i.e., the molecular weight of RT-ACE was estimated to be 104 kilo-dalton (kd) and that of RP-ACE (150 kd) on SDS-polyacrylamide gel electrophoresis. On the other hand, the enzymochemical properties of RT-ACE were very similar to those of RP-ACE, with regard to activation by NaCl, optimum pH, Km value for N*-hippuryl-His-Leu-OH hydrolysis and sensitivities to various inhibitors. Therefore, it was speculated that the portions contributing to the appearance of catalytic activity would be similar between RT-ACE and RP-ACE.

Animals↗
PubMed1986

Angiotensin I converting enzyme activity in the kidney of bullfrog (Rana catesbeiana).

In the present investigation, the occurrence of angiotensin I converting enzyme (EC 3.4.15.1; ACE) was first demonstrated in the kidney of bullfrog (Rana catesbeiana). Namely, a large amount of hydrolyzing activity toward N alpha-hippuryl-L-His-L-Leu-OH (HHL), a synthetic substrate of ACE, was detected in a 100,000 X g pellet fraction of the bullfrog kidney. The renal HHL hydrolyzing enzyme was solubilized from the 100,000 X g pellet with sodium deoxycholate. The solubilized bullfrog renal enzyme liberated H-His-Leu-OH from HHL. The enzyme activity was strongly activated by NaCl and slightly activated by cobalt sulfate in the presence of NaCl. These properties of the bullfrog renal enzyme agreed well with those of mammalian ACE previously reported. The bullfrog renal HHL hydrolyzing activity was completely inhibited by typical ACE inhibitors, ethylenediamine tetraacetic acid (10(-3) M), captopril (10(-5) M), SA-466 (10(-5) M) and MK-422 (10(-6) M). Furthermore, [Ile5]-angiotensin I converting activity was also detected in this enzyme fraction of bullfrog kidney and this converting activity was completely inhibited by MK-422 (10(-6) M). Thus, enzyme activities having characteristics of ACE were detected in the 100000 X g pellet of bullfrog kidney. We also detected [Val5, Asn9]-angiotensin I (bullfrog angiotensin I) converting activity in the bullfrog renal extract. This converting activity was also completely inhibited by MK-422 (10(-6) M).

Animals↗