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Effect of organic phosphates on methemoglobin reduction by ascorbic acid.

A Tomoda, S Matsukawa, M Takeshita

The Journal of Biological Chemistry

|December 10, 1976

Summary

Organic phosphates like ATP, 2,3-DPG, and IHP significantly accelerate methemoglobin reduction by ascorbic acid. This study quanties these effects and proposes a mechanism, highlighting 2,3-DPG

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Area of Science:

Biochemistry
Redox reactions
Enzyme kinetics

Background:

Methemoglobinemia is a condition where hemoglobin is oxidized, reducing oxygen transport.
Ascorbic acid is a known reducing agent for methemoglobin.
Organic phosphates influence hemoglobin's properties and interactions.

Purpose of the Study:

To investigate the effect of organic phosphates (ATP, 2,3-DPG, IHP) on the rate of methemoglobin reduction by ascorbic acid.
To determine the stoichiometry and kinetics of this reaction under various conditions.
To elucidate the mechanism of acceleration and the binding affinities of these phosphates to methemoglobin.

Main Methods:

Spectrophotometric determination of methemoglobin and ascorbic acid concentrations.
Kinetic analysis of methemoglobin reduction rates at different pH and organic phosphate concentrations.
Calculation of dissociation constants for organic phosphates binding to methemoglobin.

Main Results:

Ascorbic acid-mediated methemoglobin reduction was accelerated by ATP (3x), 2,3-DPG (4x), and IHP (10x) at pH 7.0.
The reaction proceeded stoichiometrically in the presence of IHP.
Dissociation constants at pH 7.0 were estimated: ATP (3.3 x 10(-4) M), 2,3-DPG (2 x 10(-3) M), and IHP (8 x 10(-6) M).

Conclusions:

Organic phosphates significantly enhance the rate of methemoglobin reduction by ascorbic acid.
The binding affinity of organic phosphates to methemoglobin correlates with their acceleration effect.
These findings provide insights into the physiological role of 2,3-DPG in modulating methemoglobin levels in red blood cells.

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