Reactions between nitric oxide and haemoglobin under physiological conditions

The tenet of high-affinity nitric oxide (NO) binding to a haemoglobin (Hb) has shaped our view of haem proteins and of small diffusible signaling molecules. Specifically, NO binds rapidly to haem iron in Hb (k approximate to 10(7) M-1 s(-1)) (refs 1, 2) and once bound, the NO activity is largely irretrievable (K-d approximate to 10(-5) s(-1)) (refs 3-10); the binding is purportedly so tight as to be unaffected by O-2 or CO. However, these general principles do not consider the allosteric state of Hb or the nature of the allosteric effector,and they mostly derive from the functional behaviour of fully nitrosylated Hb, whereas Hb is only partially nitrosylated in vivo(11-16). Here we show that oxygen drives the conversion of nitrosylhaemoglobin in the 'tense' T (or partially nitrosylated, deoxy) structure to S-nitrosohaemoglobin in the 'relaxed' R (or ligand-bound, oxy) structure. In the absence of oxygen, nitroxyl anion(NO-) is liberated in a reaction producing methaemoglobin. The yields of both S-nitrosohaemoglobin and methaemoeglobin are dependent on the NO/Hb ratio. These newly discovered reactions elucidate mechanisms underlying NO function in the respiratory cycle, and provide insight into the aetiology of S-nitrosothiols, methaemoglobin and its related valency hybrids. Mechanistic re-examination of NO interactions with other haem proteins containing allosteric-site thiols may be warranted.