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Nuclear magnetic resonance studies of equilibrated and reconstituted forms of hemoglobin

Thesis/Dissertation ·
OSTI ID:6407560

High resolution proton nuclear magnetic resonance (/sup 1/H NMR) spectroscopy is used to investigate the structure-function relationships and dynamic properties of human adult hemoglobin (Hb A). Several resonances arising from the heme and the bound ligand are located and assigned. The /sup 1/H NMR spectra of ferric low spin complexes of Hb A and its ..cap alpha.. and ..beta.. subunits are studied in detail. Differences in the position of hyperfine shifted resonances of the subunits comprising the isolated and tetrameric Hb forms of the met-azido complex reflect alteration of the spin state. This change in the spin state stems from axial perturbation of the hemes of specific subunits. /sup 1/H NMR study of the reaction of apo-Hb with heme reveals that the holoprotein is initially formed as a mixture of slowly interconverting isomers; each subunit possesses two possible orientations differing by a 180/sup 0/ rotation about the ..cap alpha..-..gamma.. meso axis, only one corresponding to the native heme orientation. The reaction rates for the equilibration of the heme reorientation reaction differ dramatically for met-aquo and met-azido derivatives. As a result of experiments involving isotope labeled hemins and kinetic control of the rotational disorder in Hb, resonances arising from the reversed heme are unambiguously assigned.

Research Organization:
California Univ., Davis (USA)
OSTI ID:
6407560
Country of Publication:
United States
Language:
English