L Uotila

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

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Publications (23)54.22 Total impact

  • L Uotila, M Koivusalo
    Advances in Experimental Medicine and Biology 02/1997; 414:365-71. · 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
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    ABSTRACT: Vitamin K functions in animal cells as the cofactor of the enzyme vitamin K-dependent carboxylase which catalyzes the post-translational formation of gamma-carboxyglutamyl (Gla) residues in specific vitamin K-dependent proteins. These proteins include four blood coagulation factors (prothrombin and Factors VII, IX and X), other plasma proteins (protein C, protein S and protein Z), two proteins from bone (osteocalcin or bone Gla-protein and matrix Gla-protein), and other proteins from lung, kidney, spleen, testis, placenta and other tissues. In the proteins involved in blood coagulation the Gla residues are mandatory for the activation of the inactive proenzymes; this process occurs on phospholipid surfaces to which the proenzymes are bound via Gla residues and calcium ions. The energy needed in the carboxylation reaction is obtained from the oxidation of vitamin K hydroquinone to 2,3-epoxide of the vitamin. Specific enzymes, vitamin K epoxide reductase and vitamin K quinone reductases, catalyze consecutive reactions in which the vitamin K hydroquinone is regenerated, thus allowing continued use of the vitamin K molecule for the carboxylations. The oral anticoagulants, derivatives of 4-hydroxycoumarin and indan-1,3-dione, used as therapeutic agents in thromboembolic disease, are antagonists to vitamin K preventing the catalytic use of vitamin K in the carboxylations by irreversibly inhibiting vitamin K epoxide reductase.
    Scandinavian journal of clinical and laboratory investigation. Supplementum 02/1990; 201:109-17.
  • Source
    M Koivusalo, M Baumann, L Uotila
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    ABSTRACT: Formaldehyde dehydrogenase (EC is a widely occurring enzyme which catalyzes the oxidation of S-hydroxymethylglutathione, formed from formaldehyde and glutathione, into S-formyglutathione in the presence of NAD. We determined the amino acid sequences for 5 tryptic peptides (containing altogether 57 amino acids) from electrophoretically homogeneous rat liver formaldehyde dehydrogenase and found that they all were exactly homologous to the sequence of rat liver class III alcohol dehydrogenase (ADH-2). Formaldehyde dehydrogenase was found to be able at high pH values to catalyze the NAD-dependent oxidation of long-chain aliphatic alcohols like n-octanol and 12-hydroxydodecanoate but ethanol was used only at very high substrate concentrations and pyrazole was not inhibitory. The amino acid sequence homology and identical structural and kinetic properties indicate that formaldehyde dehydrogenase and the mammalian class III alcohol dehydrogenases are identical enzymes.
    FEBS Letters 11/1989; 257(1):105-9. · 3.34 Impact Factor
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    ABSTRACT: Two separate pools of glyoxalase II were demonstrated in rat liver mitochondria, one in the intermembrane space and the other in the matrix. The enzyme was purified from both sources by affinity chromatography on S-(carbobenzoxy)glutathione-Affi-Gel 40. From both crude and purified preparations polyacrylamide gel-electrophoresis resolved multiple forms of glyoxalase II, two from the intermembrane space and five from the matrix. Among the thioesters of glutathione tested as substrates, S-D-lactoylglutathione was hydrolyzed most efficiently by the enzymes from both sources. Significant differences were observed in the specificities between the intermembrane space and matrix enzymes with S-acetoacetylglutathione, S-acetylglutathione, S-propionylglutathione and S-succinylglutathione as substrates. Pure glyoxalase II from rat liver cytosol was chemically polymerized and used as antigen. Antibodies were raised in rabbits and the antiserum was used for comparison of the two purified mitochondrial enzymes with cytosolic glyoxalase II by immunoblotting. The enzyme purified from the intermembrane space cross-reacted with the antiserum, but the matrix glyoxalase II did not. The results give evidence for the presence in rat liver mitochondria of two species of glyoxalase II with differing characteristics. Only the enzyme from the intermembrane space appears to resemble the cytosolic glyoxalase II forms.
    Biochimica et Biophysica Acta 11/1989; 993(1):7-11. · 4.66 Impact Factor
  • Lasse Uotila, Martti Koivusalo
    Coenzymes and cofactors. Glutathione. Chemical, biochemical and medical aspects, Edited by David Dolphin, Rozanne Poulson, Olga Avramović, 01/1989: pages 517-551; John Wiley & Sons, New York.
  • Lasse Uotila
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    ABSTRACT: The steady-state kinetic mechanism of vitamin K-dependent carboxylase from calf liver has been investigated by initial-velocity measurements with varying concentrations of two carboxylase substrates and constant, nonsaturating concentrations of the other two substrates. With all combinations of the varied substrates tested linear kinetics were obtained with lines intersecting on the left side of the 1/v axis in double-reciprocal plots. Thus the carboxylase has a sequential reaction mechanism which includes the quinternary complex of the enzyme with its four substrates. A mechanism with the ordered steady-state addition of all substrates to the enzyme accords well with the results. A totally random mechanism was excluded but the alternative possibility remained that part of the substrates are added in a rapid-equilibrium random reaction. Experiments with saturating constant concentrations of sodium bicarbonate and varying concentrations of the other substrates suggest that bicarbonate (CO2) is either the first or, more probably, the last substrate bound to the enzyme.
    Archives of Biochemistry and Biophysics 08/1988; 264(1):135-43. · 3.04 Impact Factor
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    ABSTRACT: Glyoxalase II (S-(2-hydroxyacyl)glutathione hydrolase, EC, which has been regarded as a cytosolic enzyme, was also found in rat liver mitochondria. The mitochondrial fraction contained about 10-15% of the total glyoxalase II activity in liver. The actual existence of the specific mitochondrial glyoxalase II was verified by showing that all of the activity of the crude mitochondrial pellet was still present in purified mitochondria prepared in a Ficoll gradient. Subfractionation of the mitochondria by digitonin treatment showed that 56% of the activity resided in the mitochondrial matrix and 19% in the intermembrane space. Partial purification of the enzyme (420-fold) was also achieved. Statistically significant differences were found in the substrate specificities of the mitochondrial and the cytosolic glyoxalase II. Electrophoresis and isoelectric focusing of either the crude mitochondrial extract or of the purified mitochondrial glyoxalase II resolved the enzyme activity into five forms with the respective pI values of 8.1, 7.5, 7.0, 6.85 and 6.6. Three of these forms (pI values 7.0-6.6) were exclusively mitochondrial, with no counterpart in the cytosol. The relative molecular mass of the partially purified enzyme, as estimated by Superose 12 gel chromatography, was 21,000. These results give evidence for the presence of mitochondrial glyoxalase II which is different from the cytosolic enzymes in several characteristics.
    Biochimica et Biophysica Acta 07/1988; 955(1):103-10. · 4.66 Impact Factor
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    ABSTRACT: Glyoxalase II (S-(2-hydroxyacyl)glutathione hydrolase, EC was purified to homogeneity and separated into two forms (alpha, pI = 8.0; beta, pI = 7.4) from both liver and brain of wistar rats by column isoelectric focusing. These forms were also found to have different electrophoretic mobilities. No significant differences were found between the alpha and beta forms from either source in the relative molecular mass (about 24,000) or in Km values using three substrates. The temperature-inactivation profiles were also similar, the two forms being stable up to 50 degrees C. Chemical modification studies with phenylglyoxal suggest that these enzyme forms probably contain arginine residues near the active site. Inactivation of alpha and beta forms by diethylpyrocarbonate and by photooxidation with methylene blue, and protection by S-D-mandeloylglutathione, a slowly reacting substrate, suggest the presence of histidine at the active site. The alpha and beta forms show different half-life values in inactivation by histidine reagents, which may be due to a difference in the active-site structures of these enzymes. The results probably indicate distinct structures (sequences) for alpha and beta forms.
    Biochimica et Biophysica Acta 03/1987; 911(3):349-55. · 4.66 Impact Factor
  • Lasse Uotila, Martti Koivusalo
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    ABSTRACT: Red cell hemolysates from nonrelated Finns were analyzed by electrofocusing on polyacrylamide gel, and formaldehyde dehydrogenase (EC was located by an activity-staining method. Three forms of the enzyme were constantly found for all the individuals studied but no variants were observed in this population (n = 217). Human liver also had three formaldehyde dehydrogenase forms with locations identical to those of the red cell formaldehyde dehydrogenase. Population genetic studies of formaldehyde dehydrogenase can easily be performed with red cell hemolysates with the techniques described here, and there is no need to use liver biopsy samples.
    Human Heredity 02/1987; 37(2):102-6. · 1.64 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; 77:320-5. · 2.19 Impact Factor
  • L Uotila, M Koivusalo
    Methods in Enzymology 02/1981; 77:314-20. · 2.19 Impact Factor
  • Lasse Uotila, Martti Koivusalo
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    ABSTRACT: The phenotypes of red cell glyoxalase I (EC were determined by polyacrylamide-gel slab electrophoresis in a Finnish population sample of 517 unrelated persons. The gene frequencies found were 0.361 for GLO1 and 0.639 for GLO2.
    Human Heredity 02/1980; 30(4):207-10. · 1.64 Impact Factor
  • L Uotila, M Koivusalo
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    ABSTRACT: Formaldehyde dehydrogenase (EC and formate dehydrogenase (EC have been isolated in pure form from pea seeds by a rapid procedure which employs column chromatographies on 5′-AMP-Sepharose, Sephacryl S-200, and DE32 cellulose. The apparent molecular weights of formaldehyde and formate dehydrogenases are, respectively, 82,300 and 80,300 by gel chromatography, and they both consist of two similar subunits. The isoelectric point of formaldehyde dehydrogenase is 5.8 and that of formate dehydrogenase is 6.2. The purified formate dehydrogenase gave three corresponding protein and activity bands in electrophoresis and isoelectric focusing on polyacrylamide gel whereas formaldehyde dehydrogenase gave only one band. Formaldehyde dehydrogenase catalyzes the formation of S-formylglutathione from formaldehyde, and glutathione. Formate dehydrogenase can, besides formate, also use S-formylglutathione and two other formate esters as substrates. S-Formylglutathione has a lower Km value (0.45 mm) than formate (2.1 mm) but the maximum velocity of S-formylglutathione is only 5.5% of that of formate. Pea extracts also contain a highly active S-formylglutathione hydrolase which has been separated from glyoxalase II (EC and partially purified. S-Formylglutathione hydrolase is apparently needed between formaldehyde and formate dehydrogenases in the metabolism of formaldehyde in pea seeds, in contrast to what was recently reported for Hansenula polymorpha, a yeast grown on methanol.
    Archives of Biochemistry and Biophysics 09/1979; 196(1):33-45. · 3.04 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 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
    European Journal of Biochemistry 05/1975; 52(3):493-503. · 3.58 Impact Factor

Publication Stats

489 Citations
54.22 Total Impact Points


  • 1974–1997
    • University of Helsinki
      • • Department of Clinical Chemistry
      • • Department of Medical Chemistry
      Helsinki, Province of Southern Finland, Finland
  • 1987
    • Università degli Studi di Perugia
      • Department of Clinical and Experimental Medicine
      Terni, Umbria, Italy