Some characteristics of urokinase released in organ culture of human kidney

Summary Plasminogen activator produced in organ culture of human kidney, i.e. in the histotypical arrangement of the tissue, was partially purified by affinity chromatography on para-aminobenzamidine coupled to Sepharose by a 6-carbon spacer, followed by gel chromatography on Sephadex G-100. The molecular weight of 2 active peaks were 27,000 and 52,000 daltons respectively. It was inhibited by DFP and by IgG antiurokinase.This study was supported by grant from the Swedish Medical Research Council (B77-17X-04523-03B).     

Type of interactions between agarose and Aspergillus niger endopolygalacturonases]

Endopolygalacturonase has been purified by binding on Sepharose 6 B gel; studies on interactions between agarose and the enzymes have shown that endopolygalacturonase chromatography on Sepharose 6 B was on ion-exchange chromatography and did not involve biospecific interactions.     

Occurrence of tetrodotoxin in the starfishAstropecten latespinosus

Summary A toxin causing paralysis was detected in the starfishAstropecten latespinosus. The toxin was purified by a method consisting of charcoal treatment and chromatography on CM-Sephadex C-25 and Bio-Rex 70. The toxin was identified as tetrodotoxin by its behavior in thin-layer chromatography and electrophoresis and its¹H-NMR spectrum.Acknowledgment. This work was in part supported by a research fund from the Ministry of Agriculture, Forestry and Fisheries of Japan.     

Purification of solubilized fucosyltransferase from lamb brain using ethylagarose gel]

The glycoprotein: fucosyl-transferase of the cerebral hemispheres, assayed with desialylated fetuin as exogenous acceptor, was the most active of the protein: glycosyltransferases tested in the brain. The addition of Titron X-100 to the membrane suspension, followed by high speed centrifugation, led to a solubilization of the enzyme. The use of hydrophobic chromatography on ethylagarose gave a good purification of this solubilized fucosyl-transferase, whose homogeneity has been shown by Ultrogel AcA 22, DEAE-cellulose chromatography and disc electrophoresis.     

Purification and some properties of hemagglutinin from the Myxomycete,Physarum polycephalum

A new hemagglutinin was isolated from the plasmodium ofPhysarum polycephalum by salting out with ammonium sulphate followed by chromatography on DE-32, DEAE-Toyopearl and hydroxyapatite. This hemagglutinin, named physarumin, was purified 1000-fold over crude extracts. The molecular weight of physarumin was determined to be 22,000 by size exclusion chromatography on Bio-Gel P-60 and 8,700 by SDS-polyacrylamide gel electrophoresis. Physarumin agglutinated rabbit, guinea pig, horse and human erythrocytes. Physarumin-induced hemagglutination was inhibited by fetuin and ₁ glycoprotein, but not by commercially available mono-and disaccharides. Hemagglutinating activity was blocked by EDTA, and was restored by adding Ca²⁺ but not by Mg²⁺.     

Isolation and characterization of biologically active lectin from Korean mistletoe, Viscum album var. Coloratum

A mistletoe lectin was isolated from water extracts of Korean mistletoe, a subspecies of Viscum album, grown on Quercus mongolica using CM-Sepharose chromatography followed by an affinity chromatography on a concanavalin A-Sepharose column. The compound proved to be a mistletoe lectin II with D-galactose and N-acetyl-D-galactosamine specificity. Matrix-assisted laser desorption time-of-flight mass spectroscopy showed it to have an average molecular mass of 62.7 kDa and to consist of two subunits of 30.6 kDa and 32.5 kDa. It was a basic protein with isoelectric points of 9.4 and 9.6 by capillary isoelectric focusing and was cytotoxic to Molt4 cell. Received 17 November 1998; received after revision 3 March 1999; accepted 3 March 1999     

Partial purification and characterization of acid phosphatase from sporulated oocysts of Eimeria tenella

Acid phosphatase of Eimeria tenella oocysts (Peak II) was purified 77-fold with a recovery of 26% using protamine sulfate precipitation, DEAE-cellulose chromatography and Sephadex G-200 gel filtration. This enzyme occurs in multiple forms as indicated by two peaks which can be separated by DEAE-cellulose chromatography and polyacrylamide gel electrophoresis. The partially purified enzyme has optimal activity at pH 4.5. With p-nitrophenyl phosphate the Km and Vmax values for (Peak II) were 25 mM and 1.57 mumol/min/mg protein, respectively. The enzyme (Peak II) is strongly inhibited by Hg++, Cu++, iodoacetamide, fluoride and molybdate. Tartrate and other divalent metal ions have no effect on enzyme activity. The partially purified Peak II phosphatase is not a glycoprotein as it is not absorbed on concanavalin-A Sepharose and its treatment with bacterial neuraminidase does not alter its elution profile through DEAE cellulose.     

Zur Trennung der C19-Steroide des Harns im Dünnschichtchromatogramm

Summary By a change of method to thin-layer chromatography, it was possible to fractionate completely the 17-ketosteroids from urine extract in one run. The chromatography was done in a horizontal manner on plates of 200×550 mm.     

Pro experimentis

Summary Simultaneous quantitative analysis of the plant hormones abscisic acid and indole-3-acetic acid from fruits ofVitis vinifera ssp. was performed by thin layer chromatography and high pressure liquid chromatography. Hormonal activity was proved in a biological test.     

Anticomplementary fraction from the poisonous secretion of the paratoid gland of the toad (Bufo marinus paracnemis Lutz)

Fractionation of the poisonous secretion of the toad Bufo marinus paracnemis Lutz, by dialysis and chromatography on QAE-Sephadex, led to the isolation of a fraction which was adsorbed to the ion exchanger. This fraction, when incubated with human serum, yielded an anticomplementary effect that was evaluated by measuring the kinetics of lytic activity on sensitized sheep red cells (classical pathway) and unsensitized rabbit cells (alternative pathway).     

Purification of the cytosol oestradiol-receptor complex from foetal guinea-pig uterus using electrofocusing on polyacrylamide plates

The 3H-oestradiol receptor complex obtained from the cytosol fraction of foetal guinea-pig uterus was purified by the following steps: column chromatography in Sephadex G-15 and Ultrogel, and by electrofocusing on polyacrylamide plates. In the final step a concentration of 15-17% of the foetal uterine oestradiol receptor protein was obtained. The isoelectric point (pl) of this receptor was determined to be 6.1-6.2.     

Trypsin in human milk

Human milk trypsin was purified by adsorption chromatography on cellulose-bound 4-aminobenzamidine; its molecular weight was about 24,000 daltons. Its concentration determined by a radioimmunoassay varies between 2.9 and 5.6 micrograms/l.     

Insect tissues, not microorganisms, produce linoleic acid in the house cricket and the American cockroach

Biosynthesis of linoleic acid, 18:2 (n-6), was unambiguously demonstrated to occur in the cockroach, Periplaneta americana, and the cricket, Acheta domesticus. Axenic tissue from both of these insect species was demonstrated by radio-gas-liquid chromatography (radio-GLC) and radio-high-performance liquid chromatography (radio-HPLC) to incorporate [1-14C]acetate and [1-14C]oleate into this essential fatty acid.     

Alkaline phosphatase binds to collagen; a hypothesis on the mechanism of extravesicular mineralization in epiphyseal cartilage

Affinity chromatography on Sepharose 4B-collagen gels was used to test the affinity of alkaline phosphatase for collagen. Results indicate that alkaline phosphatase of preosseous cartilage binds to collagen probably by electrostatic interactions, this interaction is inhibited by proteoglycan subunits. These results suggest that, in vivo, the formation of a collagen-alkaline phosphatase complex may be a step of the process leading to cartilage calcification.     

Rat liver S-adenosyl-L-homocysteine hydrolase purification by affinity column chromatography (author's transl)]

S-adenosyl-L-homocysteine hydrolase (EC 3.3.1.1) has been purified 240-fold from rat liver by affinity column chromatography on aminohexyl sepharose bound 6-mercaptopurine 9 D-riboside. The purified enzyme was homogeneous by gel electrophoresis.     

A rapid method for the purification of octopine dehydrogenase for the determination of cell metabolites

Summary Unlike other NAD⁺-dependent dehydrogenases, octopine dehydrogenase was not bound by blue Sepharose. A rapid 2-step purification procedure (gel filtration on Sephadex G-100 followed by affinity chromatography on blue Sepharose) resulted in a final preparation of octopine dehydrogenase which had a sp. act. of 65 units/mg protein and was free of contaminating NAD⁺-oxidoreductases. This preparation has been used successfully for the estimation of phospho-L-arginine, L-arginine and octopine in perchloric acid extracts.Supported by a grant from the Deutsche Forschungsgemeinschaft (Ga 241/1).     

The lectin fromViscum album L. purification by biospecific affinity chromatography

Summary A lectin fromViscum album which specifically binds to D-galactose was isolated by affinity chromatography on O-lactosyl-, O-galactosyl-polycarylamide or hydrolized sepharose 4 B. Some serological and physicochemical properties of the agglutinin are reported.     

Modulation of protein kinases and phosphoprotein phosphatases by a small acidic protein from bovine brains

A small, acidic and heat-stable protein was purified from bovine brains by column chromatography on DEAE-cellulose, Bio-Gel HTP, Affi-Gel phenothiazine and Sephadex G-75. This protein stimulates megamodulin-dependent protein kinase I from brains and phosphoprotein phosphatases from either brain or yeast. However, it inhibits cyclic AMP-dependent protein kinases from skeletal muscle.     

Effect of AD6 (8-monochloro-3-beta-diethylamino-ethyl-4-methyl-7-ethoxycarbonylme tho xy coumarin) on cyclic nucleotide phosphodiesterases in human platelets

The effect of AD6 (8-monochloro-3-beta-diethylamino-ethyl-4-methyl-7-ethoxycarbonylmeth oxy coumarin), an inhibitor of platelet aggregation, on cyclic nucleotide metabolism was investigated. AD6 inhibited selectively human platelet cyclic GMP phosphodiesterase, which was separated from cyclic AMP phosphodiesterase by DEAE-cellulose chromatography. Addition of AD6 to washed platelets increased cyclic GMP levels significantly in two minutes. These results could be useful in elucidating the action of AD6 on intact platelets.