Kinetic mechanism of human glutathione-dependent formaldehyde dehydrogenase

P. C. Sanghani, C. L. Stone, B. D. Ray, E. V. Pindel, Thomas Hurley, W. F. Bosron

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Formaldehyde, a major industrial chemical, is classified as a carcinogen because of its high reactivity with DNA. It is inactivated by oxidative metabolism to formate in humans by glutathione-dependent formaldehyde dehydrogenase. This NAD+-dependent enzyme belongs to the family of zinc-dependent alcohol dehydrogenases with 40 kDa subunits and is also called ADH3 or χ-ADH. The first step in the reaction involves the nonenzymatic formation of the S-(hydroxymethyl)glutathione adduct from formaldehyde and glutathione. When formaldehyde concentrations exceed that of glutathione, nonoxidizable adducts can be formed in vitro. The S-(hydroxymethyl)glutathione adduct will be predominant in vivo, since circulating glutathione concentrations are reported to be 50 times that of formaldehyde in humans. Initial velocity, product inhibition, dead-end inhibition, and equilibrium binding studies indicate that the catalytic mechanism for oxidation of S-(hydroxymethyl)glutathione and 12-hydroxydodecanoic acid (12-HDDA) with NAD+ is random bi-bi. Formation of an E·NADH·12-HDDA abortive complex was evident from equilibrium binding studies, but no substrate inhibition was seen with 12-HDDA. 12-Oxododecanoic acid (12-ODDA) exhibited substrate inhibition, which is consistent with a preferred pathway for substrate addition in the reductive reaction and formation of an abortive E·NAD+·12-ODDA complex. The random mechanism is consistent with the published three-dimensional structure of the formaldehyde dehydrogenase·NAD+ complex, which exhibits a unique semi-open coenzyme-catalytic domain conformation where substrates can bind or dissociate in any order.

Original languageEnglish
Pages (from-to)10720-10729
Number of pages10
JournalBiochemistry
Volume39
Issue number35
DOIs
StatePublished - Sep 5 2000

Fingerprint

glutathione-independent formaldehyde dehydrogenase
Glutathione
Formaldehyde
Kinetics
formic acid
NAD
Substrates
Acids
Alcohol Dehydrogenase
Industrial chemicals
Coenzymes
Carcinogens
formaldehyde dehydrogenase (glutathione)
Zinc
Catalytic Domain
Metabolism
Conformations

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sanghani, P. C., Stone, C. L., Ray, B. D., Pindel, E. V., Hurley, T., & Bosron, W. F. (2000). Kinetic mechanism of human glutathione-dependent formaldehyde dehydrogenase. Biochemistry, 39(35), 10720-10729. https://doi.org/10.1021/bi9929711

Kinetic mechanism of human glutathione-dependent formaldehyde dehydrogenase. / Sanghani, P. C.; Stone, C. L.; Ray, B. D.; Pindel, E. V.; Hurley, Thomas; Bosron, W. F.

In: Biochemistry, Vol. 39, No. 35, 05.09.2000, p. 10720-10729.

Research output: Contribution to journalArticle

Sanghani, PC, Stone, CL, Ray, BD, Pindel, EV, Hurley, T & Bosron, WF 2000, 'Kinetic mechanism of human glutathione-dependent formaldehyde dehydrogenase', Biochemistry, vol. 39, no. 35, pp. 10720-10729. https://doi.org/10.1021/bi9929711
Sanghani PC, Stone CL, Ray BD, Pindel EV, Hurley T, Bosron WF. Kinetic mechanism of human glutathione-dependent formaldehyde dehydrogenase. Biochemistry. 2000 Sep 5;39(35):10720-10729. https://doi.org/10.1021/bi9929711
Sanghani, P. C. ; Stone, C. L. ; Ray, B. D. ; Pindel, E. V. ; Hurley, Thomas ; Bosron, W. F. / Kinetic mechanism of human glutathione-dependent formaldehyde dehydrogenase. In: Biochemistry. 2000 ; Vol. 39, No. 35. pp. 10720-10729.
@article{90bae689b2594dc1acc2e2b14a4c00fc,
title = "Kinetic mechanism of human glutathione-dependent formaldehyde dehydrogenase",
abstract = "Formaldehyde, a major industrial chemical, is classified as a carcinogen because of its high reactivity with DNA. It is inactivated by oxidative metabolism to formate in humans by glutathione-dependent formaldehyde dehydrogenase. This NAD+-dependent enzyme belongs to the family of zinc-dependent alcohol dehydrogenases with 40 kDa subunits and is also called ADH3 or χ-ADH. The first step in the reaction involves the nonenzymatic formation of the S-(hydroxymethyl)glutathione adduct from formaldehyde and glutathione. When formaldehyde concentrations exceed that of glutathione, nonoxidizable adducts can be formed in vitro. The S-(hydroxymethyl)glutathione adduct will be predominant in vivo, since circulating glutathione concentrations are reported to be 50 times that of formaldehyde in humans. Initial velocity, product inhibition, dead-end inhibition, and equilibrium binding studies indicate that the catalytic mechanism for oxidation of S-(hydroxymethyl)glutathione and 12-hydroxydodecanoic acid (12-HDDA) with NAD+ is random bi-bi. Formation of an E·NADH·12-HDDA abortive complex was evident from equilibrium binding studies, but no substrate inhibition was seen with 12-HDDA. 12-Oxododecanoic acid (12-ODDA) exhibited substrate inhibition, which is consistent with a preferred pathway for substrate addition in the reductive reaction and formation of an abortive E·NAD+·12-ODDA complex. The random mechanism is consistent with the published three-dimensional structure of the formaldehyde dehydrogenase·NAD+ complex, which exhibits a unique semi-open coenzyme-catalytic domain conformation where substrates can bind or dissociate in any order.",
author = "Sanghani, {P. C.} and Stone, {C. L.} and Ray, {B. D.} and Pindel, {E. V.} and Thomas Hurley and Bosron, {W. F.}",
year = "2000",
month = "9",
day = "5",
doi = "10.1021/bi9929711",
language = "English",
volume = "39",
pages = "10720--10729",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "35",

}

TY - JOUR

T1 - Kinetic mechanism of human glutathione-dependent formaldehyde dehydrogenase

AU - Sanghani, P. C.

AU - Stone, C. L.

AU - Ray, B. D.

AU - Pindel, E. V.

AU - Hurley, Thomas

AU - Bosron, W. F.

PY - 2000/9/5

Y1 - 2000/9/5

N2 - Formaldehyde, a major industrial chemical, is classified as a carcinogen because of its high reactivity with DNA. It is inactivated by oxidative metabolism to formate in humans by glutathione-dependent formaldehyde dehydrogenase. This NAD+-dependent enzyme belongs to the family of zinc-dependent alcohol dehydrogenases with 40 kDa subunits and is also called ADH3 or χ-ADH. The first step in the reaction involves the nonenzymatic formation of the S-(hydroxymethyl)glutathione adduct from formaldehyde and glutathione. When formaldehyde concentrations exceed that of glutathione, nonoxidizable adducts can be formed in vitro. The S-(hydroxymethyl)glutathione adduct will be predominant in vivo, since circulating glutathione concentrations are reported to be 50 times that of formaldehyde in humans. Initial velocity, product inhibition, dead-end inhibition, and equilibrium binding studies indicate that the catalytic mechanism for oxidation of S-(hydroxymethyl)glutathione and 12-hydroxydodecanoic acid (12-HDDA) with NAD+ is random bi-bi. Formation of an E·NADH·12-HDDA abortive complex was evident from equilibrium binding studies, but no substrate inhibition was seen with 12-HDDA. 12-Oxododecanoic acid (12-ODDA) exhibited substrate inhibition, which is consistent with a preferred pathway for substrate addition in the reductive reaction and formation of an abortive E·NAD+·12-ODDA complex. The random mechanism is consistent with the published three-dimensional structure of the formaldehyde dehydrogenase·NAD+ complex, which exhibits a unique semi-open coenzyme-catalytic domain conformation where substrates can bind or dissociate in any order.

AB - Formaldehyde, a major industrial chemical, is classified as a carcinogen because of its high reactivity with DNA. It is inactivated by oxidative metabolism to formate in humans by glutathione-dependent formaldehyde dehydrogenase. This NAD+-dependent enzyme belongs to the family of zinc-dependent alcohol dehydrogenases with 40 kDa subunits and is also called ADH3 or χ-ADH. The first step in the reaction involves the nonenzymatic formation of the S-(hydroxymethyl)glutathione adduct from formaldehyde and glutathione. When formaldehyde concentrations exceed that of glutathione, nonoxidizable adducts can be formed in vitro. The S-(hydroxymethyl)glutathione adduct will be predominant in vivo, since circulating glutathione concentrations are reported to be 50 times that of formaldehyde in humans. Initial velocity, product inhibition, dead-end inhibition, and equilibrium binding studies indicate that the catalytic mechanism for oxidation of S-(hydroxymethyl)glutathione and 12-hydroxydodecanoic acid (12-HDDA) with NAD+ is random bi-bi. Formation of an E·NADH·12-HDDA abortive complex was evident from equilibrium binding studies, but no substrate inhibition was seen with 12-HDDA. 12-Oxododecanoic acid (12-ODDA) exhibited substrate inhibition, which is consistent with a preferred pathway for substrate addition in the reductive reaction and formation of an abortive E·NAD+·12-ODDA complex. The random mechanism is consistent with the published three-dimensional structure of the formaldehyde dehydrogenase·NAD+ complex, which exhibits a unique semi-open coenzyme-catalytic domain conformation where substrates can bind or dissociate in any order.

UR - http://www.scopus.com/inward/record.url?scp=0034609526&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034609526&partnerID=8YFLogxK

U2 - 10.1021/bi9929711

DO - 10.1021/bi9929711

M3 - Article

C2 - 10978156

AN - SCOPUS:0034609526

VL - 39

SP - 10720

EP - 10729

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 35

ER -