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

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 2O 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 2O 3.