Identical linkage and cooperativity of oxygen and carbon monoxide binding to Octopus dofleini hemocyanin.
Employment of high-precision thin-layer methods has enabled detailed functional characterization of oxygen and carbon monoxide binding for (1) the fully assembled form with 70 binding sites and (2) the isolated chains with 7 binding sites of Octopus dofleini hemocyanin. The striking difference in the cooperativities of the two ligands for the assembled decamer is revealed through an examination of the binding capacities and the partition coefficient, determined as functions of the activities of both ligands. A global analysis of the data sets supported a two-state allosteric model assuming an allosteric unit of 7. Higher level allosteric interactions were not indicated. This contrasts to results obtained for arthropod hemocyanins. Oxygen and carbon monoxide experiments performed on the isolated subunit chain confirmed the presence of functional heterogeneity reported previously [Miller, K. (1985) Biochemistry 24, 4582-4586]. The analysis shows two types of binding sites in the ratio of 4:3.
Connelly
P R
PR
Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309-0215.
Gill
S J
SJ
Miller
K I
KI
Zhou
G
G
van Holde
K E
KE
D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002248:Carbon Monoxide / Q000378:metabolism; D006433:Hemocyanins / Q000378:metabolism*; D007700:Kinetics; D008962:Models, Theoretical; D049831:Octopodiformes; D010100:Oxygen / Q000378:metabolism; D011485:Protein Binding; D013816:Thermodynamics
D002248:Carbon Monoxide; D006433:Hemocyanins; D010100:Oxygen