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Abstract:

A simple and rapid method for determining the activity of dolichol monophosphate phosphatase and dolichol diphosphate phosphatase is reported. This procedure is based on the separation of the substrates, [3H]dolichol monophosphate and [3H]dolichol diphosphate, from their reaction products, [3H]dolichol and [3H]dolichol monophosphate, by thin-layer chromatography on silica gel plastic plates. The chromatography chamber is of a nonconventional type, as it has a thin slot from which the last millimeter of the plate emerges. This special chamber allows the solvent to evaporate as it reaches the top of the plate. As a consequence, the mobile compounds tend to concentrate at the edge of the plate. The dolichol monophosphate phosphatase and dolichol diphosphate phosphatase substrates, which are more negatively charged than the corresponding reaction products, remain at the origin whereas the products migrate to the edge of the plate when appropriate solvents are used. The measurement of the radioactivity present at both the origin and the edge of the plate allows rapid estimation of the enzymatic activities. © 1981.

Registro:

Documento: Artículo
Título:Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system
Autor:Boscoboinik, D.O.; Belocopitow, E.
Filiación:Instituto de Investigaciones Bioquímicas Fundación Campomar. Obligado 2490, 1428 Buenos Aires, Argentina
Palabras clave:dolichol phosphate; phosphatase; dolichol diphosphate; enzyme assay; methodology; thin layer chromatography; Animal; Chromatography, Thin Layer; Kinetics; Microsomes, Liver; Phosphoric Monoester Hydrolases; Pyrophosphatases; Radioisotope Dilution Technique; Rats; Support, Non-U.S. Gov't; Tritium
Año:1981
Volumen:114
Número:1
Página de inicio:42
Página de fin:45
DOI: http://dx.doi.org/10.1016/0003-2697(81)90448-6
Título revista:Analytical Biochemistry
Título revista abreviado:Anal. Biochem.
ISSN:00032697
CODEN:ANBCA
CAS:dolichol phosphate, 12698-55-4, 34457-14-2; phosphatase, 9013-05-2; dolichyl-phosphatase, EC 3.1.3.51; dolichyldiphosphatase, EC 3.6.1.43; Phosphoric Monoester Hydrolases, EC 3.1.3; Pyrophosphatases, EC 3.6.1.-; Tritium, 10028-17-8
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00032697_v114_n1_p42_Boscoboinik

Referencias:

  • Wedgwood, Strominger, (1980) J. Biol. Chem, 255, pp. 1120-1123
  • Idoyaga-Vargas, Belocopitow, Mentaberry, Carminatti, (1980) FEBS Lett, 112, pp. 63-66
  • Parodi, Mordoh, Krisman, Leloir, (1969) Arch. Biochem. Biophys, 132, pp. 111-117
  • Folch, Lees, Sloane Stanley, (1957) J. Biol. Chem, 226, pp. 497-509
  • Truter, (1964) J. Chromatogr, 14, pp. 57-61
  • Libbey, Day, Reverse phase thin-layer chromatography of 2,4-dinitrophenylhydrazones of n-alkanals and n-alkan-2-ones (1964) Journal of Chromatography A, 14, pp. 275-279
  • Van Den Eijden, Continuous flow thin-layer chromatography: A new technique (1974) Analytical Biochemistry, 57, pp. 321-322

Citas:

---------- APA ----------
Boscoboinik, D.O. & Belocopitow, E. (1981) . Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system. Analytical Biochemistry, 114(1), 42-45.
http://dx.doi.org/10.1016/0003-2697(81)90448-6
---------- CHICAGO ----------
Boscoboinik, D.O., Belocopitow, E. "Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system" . Analytical Biochemistry 114, no. 1 (1981) : 42-45.
http://dx.doi.org/10.1016/0003-2697(81)90448-6
---------- MLA ----------
Boscoboinik, D.O., Belocopitow, E. "Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system" . Analytical Biochemistry, vol. 114, no. 1, 1981, pp. 42-45.
http://dx.doi.org/10.1016/0003-2697(81)90448-6
---------- VANCOUVER ----------
Boscoboinik, D.O., Belocopitow, E. Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system. Anal. Biochem. 1981;114(1):42-45.
http://dx.doi.org/10.1016/0003-2697(81)90448-6