Artículo

El editor solo permite decargar el artículo en su versión post-print desde el repositorio. Por favor, si usted posee dicha versión, enviela a
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

Properties of purified porphobilinogen deaminase (PBG-D; EC 4.3.1.8) from rat harderian gland are here presented. The enzyme behaves as a monomer of M(r) 38 ± 2 kDa and is optimally active at pH 8.0-8.2. Its activation energy, determined by an Arrhenius plot, is 76.1 kJ/mol. Initial velocity studies showed a linear progress curve for uroporphyringen I formation and a hyperbolic dependence of the initial rate on substrate concentration, indicating the existence of a sequential displacement mechanism. Apparent kinetic constants, K(m) and V(m), calculated at 37°C and pH 8.0 were 1.1 μM and 170 pmol/min mg, respectively. The pH dependence of the apparent kinetic parameters revealed the ionization of residues with pK(A)/(ES) and pK(B)/(ES) of 7.4 ± 0.1 and 8.6 ± 0.1, respectively, and a pK(E) value of 8.0 ± 0.1. Incubation of PBG-D with 5.0 mM N-ethylmaleimide and 5.0 mM 5,5'- dithiobis(2-nitrobenzoic acid) at pH 8.0 led to inhibitions of 70 and 50%, respectively. The effect of pH, as well as the effect of thiol reagents, on enzyme activity strongly suggests the involvement of cysteine residue(s) in the mechanism of uroporphyrinogen I biosynthesis, in both the catalytic reaction and the substrate binding. Rat harderian gland PBG-D activity decreased with increasing concentrations of protoporphyrin IX, reaching a 40% inhibition at the in vivo concentration of the porphyrin and 7 μM PBG. Even at saturating concentrations of substrate, inhibition by protoporphyrin was not completely reversed. So, accumulated porphyrin may act as an regulator of PBG-D activity in rat harderian gland.

Registro:

Documento: Artículo
Título:Rat harderian gland porphobilinogen deaminase: Characterization studies and regulatory action of protoporphyrin IX
Autor:Cardalda, C.A.; Juknat, A.A.; Princ, F.G.; Batlle, A.
Filiación:Ctro. de Invest. Sobre Porfirinas Y., Departamento de Quimica Biologica, Univ. de Buenos Aires Cd. Univ., 1428 Buenos Aires, Argentina
Palabras clave:Cysteine residue(s); Porphobilinogen deaminase; Porphobilinogen deaminase regulation; Protoporphyrin IX; Rat harderian gland; 5,5' dithiobis(2 nitrobenzoic acid); cysteine; n ethylmaleimide; porphobilinogen deaminase; protoporphyrin; uroporphyrin; animal tissue; article; catalysis; enzyme activity; enzyme analysis; enzyme kinetics; enzyme purification; enzyme substrate; Harder gland; male; molecular weight; nonhuman; priority journal; rat
Año:1997
Volumen:347
Número:1
Página de inicio:69
Página de fin:77
DOI: http://dx.doi.org/10.1006/abbi.1997.0310
Título revista:Archives of Biochemistry and Biophysics
Título revista abreviado:ARCH. BIOCHEM. BIOPHYS.
ISSN:00039861
CODEN:ABBIA
CAS:5,5' dithiobis(2 nitrobenzoic acid), 69-78-3; cysteine, 4371-52-2, 52-89-1, 52-90-4; n ethylmaleimide, 128-53-0; porphobilinogen deaminase, 9036-47-9, 9074-91-3; protoporphyrin, 553-12-8; uroporphyrin, 18273-06-8, 26316-36-9, 553-18-4, 607-14-7
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00039861_v347_n1_p69_Cardalda

Referencias:

  • Kennedy, G.Y., (1970) Comp. Biochem. Physiol., 36, pp. 21-36
  • Sakai, T., (1981) Arch. Histol. Jpn., 44, pp. 299-333
  • Payne, A.P., (1994) J. Anat., 185, pp. 1-49
  • Spike, R.C., Johnston, H.S., McGadey, J., Moore, M.R., Thompson, G.G., Payne, A.P., (1986) J. Anat., 145, pp. 67-77
  • Tomio, J.M., Grinstein, M., (1968) Eur. J. Biochem., 6, pp. 80-83
  • Carriere, R., (1985) Anat. Rec., 213, pp. 496-504
  • Spike, R.C., Payne, A.P., Thompson, G.G., Moore, M.R., (1990) Biochim. Biophys. Acta, 1034, pp. 1-3
  • Webb, S.M., Hoffman, R.A., Puig-Domingo, M.L., Reiter, R.J., (1992) Harderian Glands: Porphyrin Metabolism, Behavioral and Endocrine Effects, , Berlin/New York: Springer Verlag
  • Moore, M.R., Thompson, G.G., Payne, A.P., McGadey, J., (1980) Int. J. Biochem., 12, pp. 501-504
  • Lin, W.L., Nadakavukaren, M.J., (1982) Comp. Biochem. Physiol., 71, pp. 539-540
  • Elder, G.H., (1976) Essays Med. Biochem., 2, pp. 75-114
  • Jordan, P.M., Warren, M.J., (1987) FEBS Lett., 225, pp. 87-92
  • Hart, G.J., Miller, A.D., Leeper, F.G., Battersby, A.R., (1987) J. Chem. Soc. Chem. Commun., pp. 1762-1765
  • Miller, A.D., Hart, G.J., Packman, L.C., Battersby, A.R., (1988) Biochem. J., 254, pp. 915-918
  • Gellerfors, P.L., Saltzgaber-Muller, J., Douglas, M.G., (1986) Biochem. J., 240, pp. 673-677
  • Thomas, S.D., Jordan, P.M., (1986) Nucleic Acids Res., 14, pp. 6215-6226
  • Alefounder, P.A., Abell, C., Battersby, A.R., (1988) Nucleic Acids Res., 16, p. 9871
  • Raich, N., Romeo, P.H., Dubart, A., Beaupain, D., Cohen-Sola, I.M., Goossens, M., (1986) Nucleic Acids Res., 15, pp. 5955-5968
  • Beaumont, C., Porcher, C., Picat, C., Nordmann, Y., Grandchamp, B., (1989) J. Biol. Chem., 264, pp. 14829-14834
  • Sharif, A.L., Smith, A.G., Abell, C., (1989) Eur. J. Biochem., 184, pp. 353-359
  • Witty, M., Wallace-Cook, A., Albrecht, H., Spano, A., Michel, H., Shabanowitz, J., Hunt, D., Smith, A.G., (1993) Plant Physiol., 103, pp. 139-147
  • Lim, S.H., Witty, M., Wallace-Cook, A.D., Ilag, L.Y., Smith, A.G., (1994) Plant Mol. Biol., 26, pp. 863-872
  • Sancovich, H.A., Ferramola, A.M., Batlle, A.M., Del, C., Grinstein, M., (1970) Methods in Enzymology, pp. 220-222. , H. Tabor, & C.W. Tabor. New York/London: Academic Press
  • Rimington, C., (1960) Biochem. J., 75, pp. 620-623
  • Seubert, S., Seubert, A., (1989) Merck Spectr., 2, pp. 8-9
  • Lim, C.K., Rideout, J.M., Wright, D.J., (1983) Biochem. J., 211, pp. 435-438
  • Bradford, M.M., (1976) Anal. Biochem., 72, pp. 248-254
  • Williams, D.C., Morgan, C.S., McDonald, E., Battersby, A.R., (1981) Biochem. J., 193, pp. 301-310
  • Shoi, Y., Nagamine, M., Kuraki, M., Sasa, T., (1980) Biochim. Biophys. Acta, 616, pp. 300-309
  • Spano, A.J., Timko, M.P., (1991) Biochim. Biophys. Acta, 1076, pp. 29-36
  • Jones, R.M., Jordan, P.M., (1994) Biochem. J., 299, pp. 895-902
  • Juknat, A.A., Doernemann, D., Senger, H., (1994) Planta, 193, pp. 123-130
  • Anderson, P.M., Desnick, R.J., (1980) J. Biol. Chem., 255, pp. 1993-1999
  • Williams, D.C., (1984) Biochem. J., 217, pp. 675-683
  • Battersby, A.R., Fookes, C.J.R., Hart, G., Matcham, G.W.J., Pandey, P.S., (1983) J. Chem. Soc. Perkin Trans. I, pp. 3041-3047
  • Elliot, K.R., Tipton, K.F., (1974) Biochem. J., 141, pp. 789-805
  • Cleland, W.W., (1963) Biochim. Biophys. Acta, 67, pp. 104-172
  • Battersby, A.R., Fookes, C.J.R., Matcham, G.W.J., McDonald, E., Hollenstein, R., (1983) J. Chem. Soc. Perkin Trans. I, pp. 3031-3040
  • Tipton, K.F., Dixon, H.B.F., (1979) Methods Enzymol., 63, pp. 183-234
  • Hart, G.J., Miller, A.D., Battersby, A.R., (1988) Biochem. J., 252, pp. 909-912
  • Jordan, P.M., Warren, M.J., Williams, H.J., Stolowich, N.J., Roessner, C.A., Grant, S.K., Scott, A.I., (1988) FEBS Lett., 235, pp. 189-193
  • Lander, M., Pitt, A.R., Alefounder, P.R., Bardy, D., Abell, C., Battersby, A., (1991) Biochem. J., 275, pp. 447-452
  • Jordan, P.M., Woodcock, S.C., (1991) Biochem. J., 280, pp. 445-449
  • Meissner, P., Adams, P., Kirsch, R., (1993) J. Clin. Invest., 91, pp. 1436-1444

Citas:

---------- APA ----------
Cardalda, C.A., Juknat, A.A., Princ, F.G. & Batlle, A. (1997) . Rat harderian gland porphobilinogen deaminase: Characterization studies and regulatory action of protoporphyrin IX. Archives of Biochemistry and Biophysics, 347(1), 69-77.
http://dx.doi.org/10.1006/abbi.1997.0310
---------- CHICAGO ----------
Cardalda, C.A., Juknat, A.A., Princ, F.G., Batlle, A. "Rat harderian gland porphobilinogen deaminase: Characterization studies and regulatory action of protoporphyrin IX" . Archives of Biochemistry and Biophysics 347, no. 1 (1997) : 69-77.
http://dx.doi.org/10.1006/abbi.1997.0310
---------- MLA ----------
Cardalda, C.A., Juknat, A.A., Princ, F.G., Batlle, A. "Rat harderian gland porphobilinogen deaminase: Characterization studies and regulatory action of protoporphyrin IX" . Archives of Biochemistry and Biophysics, vol. 347, no. 1, 1997, pp. 69-77.
http://dx.doi.org/10.1006/abbi.1997.0310
---------- VANCOUVER ----------
Cardalda, C.A., Juknat, A.A., Princ, F.G., Batlle, A. Rat harderian gland porphobilinogen deaminase: Characterization studies and regulatory action of protoporphyrin IX. ARCH. BIOCHEM. BIOPHYS. 1997;347(1):69-77.
http://dx.doi.org/10.1006/abbi.1997.0310