NO reactions with sol-gel and solution phase samples of the ferric nitrite derivative of HbA.

Nitric Oxide : Biology and Chemistry
Camille J Roche, Joel M Friedman

Abstract

The reaction of nitric oxide (NO) with the ferric (met) nitrite derivative of human adult hemoglobin Hb is probed for both solution phase and sol-gel encapsulated populations. The evolution of both the Q band absorption spectrum and fitted populations of Hb derivatives are used to show the sequence of events occurring when NO interacts with nitrite bound to a ferric heme in Hb. The sol-gel is used to compare the evolving populations as a function of quaternary state for the starting met-nitrite populations. The redox status of intermediates is probed using the CN(-) anion to trap ferric heme species. The emergent presence of reactive NO species such as N(2)O(3) during the course of the reaction is probed using the fluorescent probe DAF-2 whereas the fluorophore Chemifluor is used as an indirect measure of the ability of the reaction to create S-nitrosothiols on glutathione. The results are consistent with the formation of a stable reactive intermediate capable of generating bioactive forms of NO. The patterns observed are consistent with a proposed mechanism whereby NO reacts with the ferric nitrite derivative to generate N(2)O(3).

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