X-Message-Number: 32605
Date: Thu, 3 Jun 2010 22:23:27 -0700 (PDT)
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Subject: carbon monoxide likely reduces glycerol toxicity
Biochem Pharmacol. 1990 Feb 15;39(4):697-705.
Oxidation of glycerol to formaldehyde by rat liver microsomes. Effects of
cytochrome P-450 inducing agents.
Winters DK, Cederbaum AI. Department of Biochemistry, Mount Sinai School of
Medicine, New York, NY 10029.
Abstract
Glycerol was shown recently to be metabolized to formaldehyde by microsomes
from chowfed control rats (Winters et al., Biochem Biophys Res Commun 153:
612-617, 1988). In the present study, experiments were carried out to
evaluate the oxidation of glycerol by microsomes isolated from rats treated
with inducers of different isozymes of cytochrome P-450. The oxidation of
glycerol to formaldehyde was increased in microsomes from rats treated with
pyrazole, ethanol or acetone relative to their respective controls, but not
after treatment with phenobarbital or 3-methylcholanthrene. This reaction
was sensitive to inhibition by carbon monoxide and was inhibited by
compounds known to be effective substrates for P-450j, e.g. aniline,
ethanol, pyrazole and 4-methylpyrazole. Treatment with pyrazole caused an
increase in Vmax for glycerol oxidation but did not affect affect the Km
(about 15 mM) for glycerol, as compared to saline controls. Evidence that
the product of glycerol metabolism is formaldehyde was provided by the
observation that this product served as a substrate for the
glutathione-dependent formaldehyde dehydrogenase, and the amount of
formaldehyde detected was identical to that detected by the Nash reaction.
By utilizing [14C]glycerol, and coupling the formaldehyde dehydrogenase
reaction to the formate dehydrogenase reaction, 14CO2 could be detected,
indicating that the formaldehyde produced was derived from the added
glycerol. These results suggest that that glycerol is not metabolically
inert when added to microsomes but serves as an effective substrate for the
cytochrome P-450j isozyme, extending the alcohol substrate specificity of
this enzyme to poly-ols. The production of formaldehyde from glycerol may
require caution since glycerol is often present in microsomal or
reconstituted systems.
PMID: 2306278
Biochem Biophys Res Commun. 1988 Jun 16;153(2):612-7.
Oxidation of glycerol to formaldehyde by rat liver microsomes.
Winters DK, Clejan LA, Cederbaum AI. Department of Biochemistry, Mount Sinai
School of Medicine, (CUNY), N.Y. 10029.
Abstract
Rat liver microsomes catalyzed the oxidation of glycerol to a Nash-reactive
material in a time- and protein-dependent manner. Omission of the glycerol
or the microsomes or any of the components of the NADPH-generating system
resulted in almost a complete loss of product formation. Apparent Km and
Vmax values for glycerol oxidation were about 18 mM and 2.5 nmol
formaldehyde per min per mg microsomal protein. Carbon monoxide inhibited
glycerol oxidation indicating a requirement for cytochrome P-450. That the
Nash-reactive material was formaldehyde was validated by a
glutathione-dependent formaldehyde dehydrogenase positive reaction. These
studies indicate that glycerol is not inert when utilized with microsomes or
reconstituted mixed function oxidase systems, and that the production of
formaldehyde from glycerol may interfere with assays of other substrates
which generate formaldehyde as product.
PMID: 3382392
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