THE 1.9-ANGSTROM STRUCTURE OF DEOXY-BETA(4) HEMOGLOBIN - ANALYSIS OF THE PARTITIONING OF QUATERNARY-ASSOCIATED AND LIGAND-INDUCED CHANGES IN TERTIARY STRUCTURE

The crystal structure of the deoxygenated form of the human hemoglobin β4 tetramer (deoxyβ4) has been determined and refined at a resolution of 1.9 Å. A detailed comparison of the quaternary structures of carbonmonoxy-β4 (COβ4) and deoxyβ4 shows that ligand binding to the β4 tetramer produces only slight movements of the subunits relative to each other. Therefore, unlike the hemoglobin α2β2 tetramer, where the transition from an unliganded T state tetramer to a liganded R state tetramer results in a large change in quaternary structure, β4 is locked in a quaternary structure that very closely resembles the R state. By comparing the high-resolution structures of T state deoxy α2β2, R state deoxyβ4 and R state COβ4, it is possible to partition the changes in β subunit tertiary structure into those that arise from changes in quaternary structure and those that result solely from ligand binding. Specifically, when viewed from the heme reference frame, comparison of the structures of T state deoxy α2β2 and R state deoxyβ4 shows that the T-to-R quaternary structure transition induces changes in β subunit tertiary structure that are approximately halfway toward the tertiary structure observed in liganded β4 and liganded α2β2. When viewed from the reference frame of the globin backbone atoms, the T-to-R quaternary structure transition induces a small rotation of the heme group and a shift of the "allosteric core" (the end of the F helix, the FG corner, the beginning of the G helix, and the heme group) away from the E helix. These movements open the ligand binding pocket and place the heme in a more symmetric position relative to the proximal histidine residue. Together, these effects work in unison to give the subunits of deoxyβ4 a tertiary structure that has high ligand affinity.