Artículo

Noriega, G.; Mattei, G.; Batlle, A.; Juknat, A.A. "Rat kidney porphobilinogen deaminase kinetics: Detection of enzyme-substrate complexes" (2002) International Journal of Biochemistry and Cell Biology. 34(10):1230-1240
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Abstract:

Background and aims: Acute intermittent porphyria (AIP) is an inherited disease resulting from a reduced activity of the enzyme porphobilinogen deaminase (PBG-D). The kidney is an important target for numerous porphyrinogenic drugs and it may contribute to the clinical manifestations of porphyric attacks. An evaluation of kidney PBG-D role in the AIP pathophysiology requires detailed information on kidney PBG-D properties, under normal conditions. Methods: Rat kidney PBG-D was purified to homogeneity and initial reaction velocities were calculated by measuring uroporphyrinogen I formation at pH 8.2 for different incubation times (0-20min) and over a wide range of substrate concentrations (0.8-66μM). Results: Purified rat kidney PBG-D is a monomeric enzyme showing only a single protein band after SDS-PAGE, Western blot and isoelectric focusing (pI 4.9). Its molecular mass is 40±2.3kDa, determined by SDS-PAGE and 39.8±2kDa by gel filtration chromatography. Rat kidney PBG-D has an unusual kinetic behaviour, exhibiting a deviation from the Michaelis-Menten hyperbola. PBG-D kinetic data required a fitting to an equation of higher degree, leading to the following apparent kinetic constants: K1=2.08±0.01μM and K2=0.102±0.003μM. Conclusion: The values of these constants fulfil the restriction 4K2≤K1 2, necessary for the occurrence of isoenzymes, interpreted in this work as enzyme-substrate intermediates. The initial reaction velocity expression here defined, correlates with an enzyme carrying only one active site but allowing, through conformational changes, the detection of at least two enzyme-substrate intermediates formed during PBG-D reaction. © 2002 Elsevier Science Ltd. All rights reserved.

Registro:

Documento: Artículo
Título:Rat kidney porphobilinogen deaminase kinetics: Detection of enzyme-substrate complexes
Autor:Noriega, G.; Mattei, G.; Batlle, A.; Juknat, A.A.
Filiación:Centro de Investigaciones sobre Porfirinas y Porfirias (CIPYP) CONICET, Departamento de Quı́mica Biológica, 1428 Buenos Aires, Argentina
Departamento de Fı́sica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, 1428 Buenos Aires, Argentina
Palabras clave:Enzyme-substrate complexes; Isoenzymes; Kinetic mechanism; Porphobilinogen deaminase; Rat kidney; porphobilinogen deaminase; isoenzyme; kidney enzyme; porphobilinogen deaminase; uroporphyrinogen; animal; article; enzyme specificity; enzymology; isoelectric focusing; kidney; kinetics; male; metabolism; polyacrylamide gel electrophoresis; rat; Western blotting; animal tissue; conformational transition; controlled study; enzyme active site; enzyme purification; enzyme substrate complex; incubation time; kidney; molecular cloning; molecular weight; nonhuman; pH; protein expression; Animals; Blotting, Western; Electrophoresis, Polyacrylamide Gel; Hydroxymethylbilane Synthase; Isoelectric Focusing; Kidney; Kinetics; Male; Rats; Substrate Specificity; Animal; Blotting, Western; Electrophoresis, Polyacrylamide Gel; Isoelectric Focusing; Kinetics; Male; Rats; Substrate Specificity; Support, Non-U.S. Gov't
Año:2002
Volumen:34
Número:10
Página de inicio:1230
Página de fin:1240
DOI: http://dx.doi.org/10.1016/S1357-2725(02)00051-1
Título revista:International Journal of Biochemistry and Cell Biology
Título revista abreviado:Int. J. Biochem. Cell Biol.
ISSN:13572725
CAS:Hydroxymethylbilane Synthase, EC 2.5.1.61; Hydroxymethylbilane Synthase, EC 4.3.1.8
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_13572725_v34_n10_p1230_Noriega

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Citas:

---------- APA ----------
Noriega, G., Mattei, G., Batlle, A. & Juknat, A.A. (2002) . Rat kidney porphobilinogen deaminase kinetics: Detection of enzyme-substrate complexes. International Journal of Biochemistry and Cell Biology, 34(10), 1230-1240.
http://dx.doi.org/10.1016/S1357-2725(02)00051-1
---------- CHICAGO ----------
Noriega, G., Mattei, G., Batlle, A., Juknat, A.A. "Rat kidney porphobilinogen deaminase kinetics: Detection of enzyme-substrate complexes" . International Journal of Biochemistry and Cell Biology 34, no. 10 (2002) : 1230-1240.
http://dx.doi.org/10.1016/S1357-2725(02)00051-1
---------- MLA ----------
Noriega, G., Mattei, G., Batlle, A., Juknat, A.A. "Rat kidney porphobilinogen deaminase kinetics: Detection of enzyme-substrate complexes" . International Journal of Biochemistry and Cell Biology, vol. 34, no. 10, 2002, pp. 1230-1240.
http://dx.doi.org/10.1016/S1357-2725(02)00051-1
---------- VANCOUVER ----------
Noriega, G., Mattei, G., Batlle, A., Juknat, A.A. Rat kidney porphobilinogen deaminase kinetics: Detection of enzyme-substrate complexes. Int. J. Biochem. Cell Biol. 2002;34(10):1230-1240.
http://dx.doi.org/10.1016/S1357-2725(02)00051-1