Deuterohemins III and XIII were coupled with apomyoglobin to examine the influence of the modified heme-globin contacts on the functions of the reconstituted holoproteins. Owing to the molecular symmetry of the prosthetic groups, the resultant proteins are free from the heme orientational disorder. The coordination structures of the two reconstituted myoglobins were found to be normal and closely similar to each other. The equilibrium ligand bindings also resembled with each other in both ferric and ferrous states. The results demonstrate that the different local heme-globin contacts affect the structure and function of the reconstituted myoglobins only slightly. The results therefore suggest that the two asymmetric deuteroheme IX isomers, which are inverted about the alpha, gamma-meso carbon axis of the heme, also exhibit very similar functions in myoglobin.