Mechanism of inhibition of aldose reductase by menadione (vitamin K3)

Aruni Bhatnagar, Si Qi Liu, J. Mark Petrash, Satish Srivastava

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Incubation of human placental aldose reductase (EC 1.1.1.21) with menadione (0.5-3.0 mM) resulted in time-dependent loss of the catalytic activity of the enzyme. Kinetic analysis of the data suggests that the inactivation process follows a single apparent rate constant that displays hyperbolic dependence on menadione concentration, indicating that menadione forms a kinetically significant, dissociable complex with the enzyme before the formation of an inactive enzyme-menadione complex. The inactivation of the enzyme with menadione was reversed upon dialysis of the inactivated enzyme against buffer containing 10 mM dithiothreitol suggesting that menadione reacts with enzyme sulfhydryl residue(s). Inactivation of the enzyme was significantly prevented by dithiothreitol (5 mM), NADPH (0.1 mM), and DL-glyceraldehyde (10 mM). Correlation of the fractional remaining activity with the extent of modification indicates that loss of catalytic activity corresponds to the modification of a single amino acid residue of the enzyme protein. Recombinant human aldose reductase, obtained by overexpression in Escherichia coli, and aldose reductase in which Cys-80 or Cys-303 was replaced by serine were also inactivated by menadione. However, enzyme in which Cys-298 was replaced by serine was insensitive to menadione. On the basis of these observations, it is suggested that menadione forms a thiodione-like adduct with Cys-298, leading to inactivation of the enzyme.

Original languageEnglish (US)
Pages (from-to)917-921
Number of pages5
JournalMolecular Pharmacology
Volume42
Issue number5
StatePublished - Nov 1992

Fingerprint

Vitamin K 3
Aldehyde Reductase
Enzymes
Dithiothreitol
Serine
Glyceraldehyde
NADP
Dialysis
Buffers
Escherichia coli

ASJC Scopus subject areas

  • Pharmacology

Cite this

Bhatnagar, A., Liu, S. Q., Petrash, J. M., & Srivastava, S. (1992). Mechanism of inhibition of aldose reductase by menadione (vitamin K3). Molecular Pharmacology, 42(5), 917-921.

Mechanism of inhibition of aldose reductase by menadione (vitamin K3). / Bhatnagar, Aruni; Liu, Si Qi; Petrash, J. Mark; Srivastava, Satish.

In: Molecular Pharmacology, Vol. 42, No. 5, 11.1992, p. 917-921.

Research output: Contribution to journalArticle

Bhatnagar, A, Liu, SQ, Petrash, JM & Srivastava, S 1992, 'Mechanism of inhibition of aldose reductase by menadione (vitamin K3)', Molecular Pharmacology, vol. 42, no. 5, pp. 917-921.
Bhatnagar A, Liu SQ, Petrash JM, Srivastava S. Mechanism of inhibition of aldose reductase by menadione (vitamin K3). Molecular Pharmacology. 1992 Nov;42(5):917-921.
Bhatnagar, Aruni ; Liu, Si Qi ; Petrash, J. Mark ; Srivastava, Satish. / Mechanism of inhibition of aldose reductase by menadione (vitamin K3). In: Molecular Pharmacology. 1992 ; Vol. 42, No. 5. pp. 917-921.
@article{358832b0e2374309bed3afa2f875fba6,
title = "Mechanism of inhibition of aldose reductase by menadione (vitamin K3)",
abstract = "Incubation of human placental aldose reductase (EC 1.1.1.21) with menadione (0.5-3.0 mM) resulted in time-dependent loss of the catalytic activity of the enzyme. Kinetic analysis of the data suggests that the inactivation process follows a single apparent rate constant that displays hyperbolic dependence on menadione concentration, indicating that menadione forms a kinetically significant, dissociable complex with the enzyme before the formation of an inactive enzyme-menadione complex. The inactivation of the enzyme with menadione was reversed upon dialysis of the inactivated enzyme against buffer containing 10 mM dithiothreitol suggesting that menadione reacts with enzyme sulfhydryl residue(s). Inactivation of the enzyme was significantly prevented by dithiothreitol (5 mM), NADPH (0.1 mM), and DL-glyceraldehyde (10 mM). Correlation of the fractional remaining activity with the extent of modification indicates that loss of catalytic activity corresponds to the modification of a single amino acid residue of the enzyme protein. Recombinant human aldose reductase, obtained by overexpression in Escherichia coli, and aldose reductase in which Cys-80 or Cys-303 was replaced by serine were also inactivated by menadione. However, enzyme in which Cys-298 was replaced by serine was insensitive to menadione. On the basis of these observations, it is suggested that menadione forms a thiodione-like adduct with Cys-298, leading to inactivation of the enzyme.",
author = "Aruni Bhatnagar and Liu, {Si Qi} and Petrash, {J. Mark} and Satish Srivastava",
year = "1992",
month = "11",
language = "English (US)",
volume = "42",
pages = "917--921",
journal = "Molecular Pharmacology",
issn = "0026-895X",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "5",

}

TY - JOUR

T1 - Mechanism of inhibition of aldose reductase by menadione (vitamin K3)

AU - Bhatnagar, Aruni

AU - Liu, Si Qi

AU - Petrash, J. Mark

AU - Srivastava, Satish

PY - 1992/11

Y1 - 1992/11

N2 - Incubation of human placental aldose reductase (EC 1.1.1.21) with menadione (0.5-3.0 mM) resulted in time-dependent loss of the catalytic activity of the enzyme. Kinetic analysis of the data suggests that the inactivation process follows a single apparent rate constant that displays hyperbolic dependence on menadione concentration, indicating that menadione forms a kinetically significant, dissociable complex with the enzyme before the formation of an inactive enzyme-menadione complex. The inactivation of the enzyme with menadione was reversed upon dialysis of the inactivated enzyme against buffer containing 10 mM dithiothreitol suggesting that menadione reacts with enzyme sulfhydryl residue(s). Inactivation of the enzyme was significantly prevented by dithiothreitol (5 mM), NADPH (0.1 mM), and DL-glyceraldehyde (10 mM). Correlation of the fractional remaining activity with the extent of modification indicates that loss of catalytic activity corresponds to the modification of a single amino acid residue of the enzyme protein. Recombinant human aldose reductase, obtained by overexpression in Escherichia coli, and aldose reductase in which Cys-80 or Cys-303 was replaced by serine were also inactivated by menadione. However, enzyme in which Cys-298 was replaced by serine was insensitive to menadione. On the basis of these observations, it is suggested that menadione forms a thiodione-like adduct with Cys-298, leading to inactivation of the enzyme.

AB - Incubation of human placental aldose reductase (EC 1.1.1.21) with menadione (0.5-3.0 mM) resulted in time-dependent loss of the catalytic activity of the enzyme. Kinetic analysis of the data suggests that the inactivation process follows a single apparent rate constant that displays hyperbolic dependence on menadione concentration, indicating that menadione forms a kinetically significant, dissociable complex with the enzyme before the formation of an inactive enzyme-menadione complex. The inactivation of the enzyme with menadione was reversed upon dialysis of the inactivated enzyme against buffer containing 10 mM dithiothreitol suggesting that menadione reacts with enzyme sulfhydryl residue(s). Inactivation of the enzyme was significantly prevented by dithiothreitol (5 mM), NADPH (0.1 mM), and DL-glyceraldehyde (10 mM). Correlation of the fractional remaining activity with the extent of modification indicates that loss of catalytic activity corresponds to the modification of a single amino acid residue of the enzyme protein. Recombinant human aldose reductase, obtained by overexpression in Escherichia coli, and aldose reductase in which Cys-80 or Cys-303 was replaced by serine were also inactivated by menadione. However, enzyme in which Cys-298 was replaced by serine was insensitive to menadione. On the basis of these observations, it is suggested that menadione forms a thiodione-like adduct with Cys-298, leading to inactivation of the enzyme.

UR - http://www.scopus.com/inward/record.url?scp=0027055224&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027055224&partnerID=8YFLogxK

M3 - Article

VL - 42

SP - 917

EP - 921

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 0026-895X

IS - 5

ER -