Nanosecond absorption spectroscopy of hemoglobin: elementary processes in kinetic cooperativity.
A nanosecond absorption spectrometer has been used to measure the optical spectra of hemoglobin between 3 ns and 100 ms after photolysis of the CO complex. The data from a single experiment comprise a surface, defined by the time-ordered set of 50-100 spectra. Singular value decomposition is used to represent the observed spectra in terms of a minimal set of basis spectra and the time course of their amplitudes. Both CO rebinding and conformational changes are found to be multiphasic. Prior to the quaternary structural change, two relaxations are observed that are assigned to geminate recombination followed by a tertiary structural change. These relaxations are interpreted in terms of a kinetic model that points out their potential role in kinetic cooperativity. The rapid escape of CO from the heme pocket compared with the rate of rebinding observed for both R and T quaternary states shows that the quaternary structure controls the overall dissociation rate by changing the rate at which the Fe--CO bond is broken. A comparable description of the control of the overall association rates must await a more complete experimental description of the kinetics of the quaternary T state.
D000494:Allosteric Regulation; D002263:Carboxyhemoglobin; D006441:Hemoglobin A; D006801:Humans; D007700:Kinetics; D011487:Protein Conformation; D013057:Spectrum Analysis; D013329:Structure-Activity Relationship
D006441:Hemoglobin A; D002263:Carboxyhemoglobin