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Abstract:

A simple mathematical model for studying mechanism-based inhibitors (MBIs) is presented. The mathematical equations are deduced for an experimental protocol consisting of a first incubation of the enzyme in the presence of MBI followed by a washing protocol to eliminate free MBI. Finally enzyme activity (initial velocity) is measured with specific substrate. The representation of the final equation obtained is a straight line, and the MBI-specific association constant of velocity (k) can be calculated from its slope. The mathematical model was then challenged with the effect of 18-ethynyl-11-deoxycorticosterone (18-EtDOC) as an MBI on aldosterone biosynthesis from 11-deoxycorticosterone (DOC) in rat adrenal mitochondria. The last step of the mitochondrial biosynthesis of aldosterone consists of the conversion of DOC into corticosterone (B) or 18-hydroxy-11-deoxycorticosterone (18-OHDOC), and both steroids can then be transformed into aldosterone. The k (mM-1·min-1) values obtained for 18-EtDOC were: 451 ± 36 for DOC to aldosterone; 177 ± 16 for B to aldosterone; 175 ± 15 for 18-OHDOC to aldosterone; and 2.7 ± 0.2 for DOC to B. These results show that this MBI practically does not affect the metabolism of DOC to B in our enzyme preparation and that conversions of B and 18-OHDOC into aldosterone are catalyzed by the same enzyme.

Registro:

Documento: Artículo
Título:Inhibition of aldosterone production in rat adrenal mitochondria by 18-ethynyl-11-deoxycorticosterone: A simple model for kinetic interpretation of mechanism-based inhibitors
Autor:Matkovic, L.; Gomez-Sanchez, C.E.; Cozza, E.N.
Filiación:Department of Biological Chemistry, University of Buenos Aires, PRHOM-CONICET, 1428-Buenos Aires, Argentina
Division of Endocrinology, University of Mississippi Medical Center, Jackson, MS 39612, United States
G.V. (Sonny) Montgomery VA Medical Center, Jackson, MS 39216, United States
Palabras clave:18-ethynyl-11-deoxycorticosterone; Adrenal gland; Aldosterone; Mechanism-based inhibitors; Suicide inhibitors; 18 ethynyl 11 deoxycorticosterone; aldosterone; deoxycorticosterone; suicide substrate; unclassified drug; adrenal gland; animal cell; animal tissue; article; controlled study; enzyme activity; enzyme mechanism; enzyme substrate; mathematical model; mitochondrion; nonhuman; rat; Animalia
Año:2001
Volumen:129
Número:3
Página de inicio:383
Página de fin:390
DOI: http://dx.doi.org/10.1093/oxfordjournals.jbchem.a002868
Título revista:Journal of Biochemistry
Título revista abreviado:J. Biochem.
ISSN:0021924X
CODEN:JOBIA
CAS:aldosterone, 52-39-1, 6251-69-0; deoxycorticosterone, 64-85-7
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0021924X_v129_n3_p383_Matkovic

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Citas:

---------- APA ----------
Matkovic, L., Gomez-Sanchez, C.E. & Cozza, E.N. (2001) . Inhibition of aldosterone production in rat adrenal mitochondria by 18-ethynyl-11-deoxycorticosterone: A simple model for kinetic interpretation of mechanism-based inhibitors. Journal of Biochemistry, 129(3), 383-390.
http://dx.doi.org/10.1093/oxfordjournals.jbchem.a002868
---------- CHICAGO ----------
Matkovic, L., Gomez-Sanchez, C.E., Cozza, E.N. "Inhibition of aldosterone production in rat adrenal mitochondria by 18-ethynyl-11-deoxycorticosterone: A simple model for kinetic interpretation of mechanism-based inhibitors" . Journal of Biochemistry 129, no. 3 (2001) : 383-390.
http://dx.doi.org/10.1093/oxfordjournals.jbchem.a002868
---------- MLA ----------
Matkovic, L., Gomez-Sanchez, C.E., Cozza, E.N. "Inhibition of aldosterone production in rat adrenal mitochondria by 18-ethynyl-11-deoxycorticosterone: A simple model for kinetic interpretation of mechanism-based inhibitors" . Journal of Biochemistry, vol. 129, no. 3, 2001, pp. 383-390.
http://dx.doi.org/10.1093/oxfordjournals.jbchem.a002868
---------- VANCOUVER ----------
Matkovic, L., Gomez-Sanchez, C.E., Cozza, E.N. Inhibition of aldosterone production in rat adrenal mitochondria by 18-ethynyl-11-deoxycorticosterone: A simple model for kinetic interpretation of mechanism-based inhibitors. J. Biochem. 2001;129(3):383-390.
http://dx.doi.org/10.1093/oxfordjournals.jbchem.a002868