Free radicals and excited species in the metabolism of indole-3-acetic acid and its ethyl ester by horseradish peroxidase and by neutrophils.

Abstract

The peroxidative metabolization of indole-3-acetic acid, a biologically important process, has been followed by EPR spectroscopy with the aim of obtaining information on the mechanism of generation of electronically excited species. The skatole-3-methylene radical detected during oxidation by horseradish peroxidase, does not appear to be involved in a major oxygen consuming process or in the generation of singlet oxygen. The chemiluminescence spectrum exhibits several maxima, which are also observed when the ethyl ester of indole-3-acetic acid is metabolized by horseradish peroxidase or by myeloperoxidase in neutrophils. When the ester is metabolically activated in either of these systems, the EPR spectrum indicates a tertiary carbon-centered radical. This radical centered on the carbon in the 3-position participates in a chemiexcitation/emissive route. Within the cell, this emissive process is responsible for a large part of the oxygen consumed. Some of the emitters originate in the cleavage of the 2,3 double bond. The ester, which is capable of penetrating into the cells, also emits with other myeloperoxidase-containing cells. This compound may have useful applications as an intracellular chemiluminescent probe for the presence of myeloperoxidase.