Oxygen-sensing mechanism of HemAT from Bacillus subtilis: a resonance Raman spectroscopic study.

Resonance Raman (RR) evidence for structural linkage between the distal side of heme pocket and the signaling domain of an oxygen sensing hemoprotein, HemAT-Bs, is reported. The band-fitting analyses of the RR spectra in the Fe-O2 stretching (nuFe-O2) region revealed the presence of three conformers with nuFe-O2 at 554, 566, and 572 cm-1, which reflect different H-bond strengths on the bound O2 molecule. While recent X-ray analysis for CN--bound HemAT-Bs suggested the importance of Thr95 and Tyr70, the species with the strongest H-bond (554 cm-1) was deleted in the T95A mutant and also by removal of the linker and signal domains; however, the Y70F mutant maintained the same three conformers. A scheme for specific O2 sensing and signaling mechanism is discussed.