Access of ligands to the ferric center in lipoxygenase-1.

A form of ferric lipoxygenase-1 has been isolated that gives an EPR spectrum that is dominated by a species of intermediate rhombicity (E/D = 0.065). This species is obtained in the presence of a number of buffers of high concentration and in the absence of fatty acid byproducts of the iron oxidation. The species is unstable over a period of one day with respect to symmetry of the iron. The EPR lineshapes of the unstable species are highly sensitive to the anionic composition of the buffer and to the addition of neutral ligands. These results suggest that newly formed ferric lipoxygenase has weak affinity for a number of ligands. Affinity of charged ligands for the iron center may provide a mechanism for charge compensation as the iron center alternates between ferric and ferrous in the catalytic cycle. We use spectral simulation to evaluate quantitatively the interaction of the ferric center with ligands and also show that a transition in the middle Kramers doublet makes a significant contribution to the EPR spectrum of the more rhombic species.