Characterization of the glutathione binding site of aldose reductase

Kota Ramana, Bharat L. Dixit, Sanjay Srivastava, Aruni Bhatnagar, Ganesaratnam K. Balendiran, Stanley Watowich, J. Mark Petrash, Satish Srivastava

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Despite extensive investigations, the physiological role of the polyol pathway enzyme-aldose reductase (AR) remains obscure. While the enzyme reduces glucose in vivo and in vitro, kinetic and structural studies indicate inefficient carbohydrate binding to the active site of the enzyme. The active site is lined by hydrophobic residues and appears more compatible with the binding of medium- to long-chain aliphatic aldehydes or hydrophobic aromatic aldehydes. In addition, our recent studies show that glutathione (GS) conjugates are also reduced efficiently by the enzyme. For instance, the GS conjugate of acrolein is reduced with a catalytic efficiency 1000-fold higher than the parent aldehyde, indicating specific recognition of glutathione by the active site residues of AR. An increase in the catalytic efficiency upon glutathiolation was also observed with trans-2-nonenal, trans-2-hexenal and trans, trans-2,4-decadienal, establishing that enhancement of catalytic efficiency was specifically due to the glutathione backbone and not specific to the aldehyde. Structure-activity relationships with substitution or deletion of amino acids of GSH indicated specific interactions of the active site with γ-Glu1 and Cys of GSH. Molecular modeling revealed that the glutathione-propanal conjugate could bind in two distinct orientations. In orientation 1, γ-Glu1 of the conjugate interacts with Trp20, Lys21 and Val47, and Gly3 interacts with Ser302 and Leu301, whereas in orientation 2, the molecule is inverted with γ-Glu1 interacting with Ser302, and Leu301. Taken together, these data suggest that glutathiolation of aldehydes enhances their compatibility with the AR active site, which may be of physiological significance in detoxification of endogenous and xenobiotic aldehydes.

Original languageEnglish (US)
Pages (from-to)537-548
Number of pages12
JournalChemico-Biological Interactions
Volume130-132
DOIs
StatePublished - Jan 30 2001

Fingerprint

Aldehyde Reductase
Aldehydes
Glutathione
Binding Sites
Catalytic Domain
Enzymes
Acrolein
Detoxification
Molecular modeling
Xenobiotics
Structure-Activity Relationship
Substitution reactions
Carbohydrates
Amino Acids
Glucose
Molecules
Kinetics

Keywords

  • Aldehydes
  • Aldose reductase
  • Glutathione
  • Regulation

ASJC Scopus subject areas

  • Toxicology

Cite this

Ramana, K., Dixit, B. L., Srivastava, S., Bhatnagar, A., Balendiran, G. K., Watowich, S., ... Srivastava, S. (2001). Characterization of the glutathione binding site of aldose reductase. Chemico-Biological Interactions, 130-132, 537-548. https://doi.org/10.1016/S0009-2797(00)00297-0

Characterization of the glutathione binding site of aldose reductase. / Ramana, Kota; Dixit, Bharat L.; Srivastava, Sanjay; Bhatnagar, Aruni; Balendiran, Ganesaratnam K.; Watowich, Stanley; Petrash, J. Mark; Srivastava, Satish.

In: Chemico-Biological Interactions, Vol. 130-132, 30.01.2001, p. 537-548.

Research output: Contribution to journalArticle

Ramana, K, Dixit, BL, Srivastava, S, Bhatnagar, A, Balendiran, GK, Watowich, S, Petrash, JM & Srivastava, S 2001, 'Characterization of the glutathione binding site of aldose reductase', Chemico-Biological Interactions, vol. 130-132, pp. 537-548. https://doi.org/10.1016/S0009-2797(00)00297-0
Ramana, Kota ; Dixit, Bharat L. ; Srivastava, Sanjay ; Bhatnagar, Aruni ; Balendiran, Ganesaratnam K. ; Watowich, Stanley ; Petrash, J. Mark ; Srivastava, Satish. / Characterization of the glutathione binding site of aldose reductase. In: Chemico-Biological Interactions. 2001 ; Vol. 130-132. pp. 537-548.
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