L Uotila

University of Helsinki, Helsinki, Province of Southern Finland, Finland

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Publications (11)20.58 Total impact

  • L Uotila · M Koivusalo
    Advances in Experimental Medicine and Biology 02/1997; 414:365-71. DOI:10.1007/978-1-4615-5871-2_42 · 2.01 Impact Factor
  • M Koivusalo · R Lapatto · L Uotila
    Advances in Experimental Medicine and Biology 02/1995; 372:427-33. · 2.01 Impact Factor
  • M Koivusalo · L Uotila
    Advances in Experimental Medicine and Biology 02/1993; 328:465-74. · 2.01 Impact Factor
  • M Koivusalo · L Uotila
    Advances in Experimental Medicine and Biology 02/1991; 284:305-13. · 2.01 Impact Factor
  • L Uotila · M Koivusalo
    Progress in clinical and biological research 02/1987; 232:165-77.
  • M Koivusalo · T Koivula · L Uotila
    Progress in clinical and biological research 02/1982; 114:155-68.
  • L Uotila · M Koivusalo
    Methods in Enzymology 02/1981; 77:314-20. · 2.19 Impact Factor
  • L Uotila · M Koivusalo
    Methods in Enzymology 02/1981; 77:320-5. DOI:10.1016/S0076-6879(81)77045-9 · 2.19 Impact Factor
  • L Uotila · M Koivusalo
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    ABSTRACT: Methods have been devised for the separation of the isoenzymes of glyoxalase I(S-lactoylglutathione methylglyoxal-lyase (isomerizing), EC 4.4.1.5) from human red blood cells by electrophoresis and electrofocusing on polyacrylamide gel slabs. Three different staining methods were used for the location of the enzyme. Three electrophoretic phenotypes of the enzyme were resolved, the fast and slow types with one band and the intermediate type with three glyoxalase I activity bands. In gel electrofocusing (pH gradient 3.5-9.5) two glyoxalase I activity bands were found for all electrophoretic types. In electrofocusing on gel with a narrow pH gradient, at least four separate enzyme components were resolved for the fast and slow electrophoretic types and at least six components for the intermediate type. The phenotypes could be distinguished correspondingly to the electrophoretic results. Preparative separation of the isoenzymes were achieved by ion exchange chromatography on DEAE-Sephacel but gel chromatography on Sephadex G-100 gave the same elution volume for all enzyme phenotypes. This corresponds to an apparent molecular weight of about 47 000.
    Acta chemica Scandinavica. Series B: Organic chemistry and biochemistry 02/1979; 34(1):63-8.
  • L Uotila · M Koivusalo
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    ABSTRACT: Glyoxalase I was obtained in electrophoretically pure form from sheep liver by a procedure which includes ammonium sulfate and poly(ethylene glycol) fractionations, and column chromatographies on hydroxypatite, Cibacron Blue Sephadex G 100 and diethylaminoethyl (DEAE) cellulose. The specific activity of the homogeneous preparations is about 4000 units/mg of protein (25°C). Three separate peaks of activity were obtained in the last column on DEAE cellulose (DE 32). No signs of heterogeneity were seen in the previous steps. Purified, but not crude preparations, gave 2 activity peaks on the disc gel electrophoresis. The isoelectric point of metal free apoglyoxalase I is 5.0 by isoelectric focusing. The apparent molecular weight of glyoxalase I is 45,900 from gel chromatography. From dodecylsulfate gel electrophoresis, the enzyme has a subunit molecular weight of 21,000. In addition to methylglyoxal, phenylglyoxal and kethoxal are good substrates of sheep liver glyoxalase I. Hydroxypyruvaldehyde and glyoxal react more slowly. Catalytically inactive apoenzyme of sheep liver glyoxalase I was prepared by dialysis against ethylenediaminetetraacetate (EDTA) and Chelex 100. All of the activity is restored by Mg2+ ions; Zn2+, Mn2+, Co2+, Ni2+ and Ca2+ ions, in decreasing order of maximum velocity, give partial reactivation. Of these metals, Mg2+ has the highest (2.8 mM), and Zn2+ the lowest (0.007 mM) apparent half saturation concentration, when assayed in 80 mM N 2 hydroxyethylpiperazine N' 2 ethanesulfonate (Hepes) buffer pH 6.8. Several chelating agents are inhibitors of sheep liver glyoxalase I. In all cases Mg2+ reverses the inhibition after a short incubation time.
    European Journal of Biochemistry 05/1975; 52(3):493-503. · 3.58 Impact Factor
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    L Uotila · M Koivusalo
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    ABSTRACT: S-Formylglutathione hydrolase, a new glutathione thiol esterase from human liver, has been purified to homogeneity according to disc electrophoretic and ultracentrifugal criteria. The final preparation catalyzed the hydrolysis of 4,100 μmoles of S-formylglutathione per min/mg of protein at 25°C and represented 2,350-fold purification over 23,000 x g supernatant of liver homogenate. In addition to S-formylglutathione, the enzyme catalyzed the hydrolysis of S-acetylglutathione 200 times slower but had no activity with S-lactylglutathione. Th elution volume of the enzyme in a gel filtration column gave an apparent molecular weight of 52,500 and a diffusion coefficient of 6.57 x 10-7 cm2 sec-1. The enzyme had a sedimentation coefficient of 4.24 S as judged from sedimentation velocity in analytical ultracentrifuge. From a sedimentation equilibrium experiment, the molecular weight of the enzyme is 55,500. Dodecyl sulfate gel electrophoresis indicated a subunit molecular weight of 30,000. Thus, the enzyme probably consists of 2 subunits. The isoelectric point of the enzyme obtained from electrofocusing experiments was 5.41. The enzyme apparently contains a reactive -SH group and purification was possible only in the presence of thiols. Activity was rapidly lost when the thiols were removed, but dithiothreitol could regenerate part of the activity. Several types of -SH reagents were inhibitory. Dithiothreitol could completely reverse the effect of mercaptide forming and oxidizing agents. Amino group reagents also rapidly inactivate the enzyme. In addition, ascorbate and folate were inhibitory. Chelating agents and organophosphates did not inhibit the enzyme.
    Journal of Biological Chemistry 01/1975; 249(23):7664-72. · 4.57 Impact Factor