The reaction of Xenopus hemoglobin with oxygen and carbon monoxide has been reinvestigated over the pH range K-6.0, in the absence and presence of organic phosphates (2,3-diphosphoglycerate or inositol hexakisphosphate), to establish if the tetramer can be stabilized in a T-quaternary state by protons and polyphosphate; the equilibrium and kinetic data indicate that Xenopus hemoglobin does exhibit a Root effect.These new results are discussed with reference to those reported by Bridges et al. [(1985) Resp. Physiol. 61, 125-1361 on Xenopus blood and, more generally, to the molecular definition and the structural basis of the Root effect as an extreme form of the Bohr effect.