Conformational dependence of hemoglobin reactivity under high viscosity conditions: the role of solvent slaved dynamics.
The concept of protein dynamic states is introduced. This concept is based on (i) protein dynamics being organized hierarchically with respect to solvent slaving and (ii) which tier of dynamics is operative over the time window of a given measurement. The protein dynamic state concept is used to analyze the kinetic phases derived from the recombination of carbon monoxide to sol-gel-encapsulated human adult hemoglobin (HbA) and select recombinant mutants. The temperature-dependent measurements are made under very high viscosity conditions obtained by bathing the samples in an excess of glycerol. The results are consistent with a given tier of solvent slaved dynamics becoming operative at a time delay (with respect to the onset of the measurement) that is primarily solvent- and temperature-dependent. However, the functional consequences of the dynamics are protein- and conformation-specific. The kinetic traces from both equilibrium populations and trapped allosteric intermediates show a consistent progression that exposes the role of both conformation and hydration in the control of reactivity. Iron-zinc symmetric hybrid forms of HbA are used to show the dramatic difference between the kinetic patterns for T state alpha and beta subunits. The overall results support a model for allostery in HbA in which the ligand-binding-induced transition from the deoxy T state to the high -affinity R state proceeds through a progression of T state intermediates.
Samuni
Uri
U
Department of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Roche
Camille J
CJ
Dantsker
David
D
Friedman
Joel M
JM
D000495:Allosteric Site; D002248:Carbon Monoxide / Q000737:chemistry; D005990:Glycerol / Q000737:chemistry; D006454:Hemoglobins / Q000737:chemistry*; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008968:Molecular Conformation; D009154:Mutation; D044367:Phase Transition; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry*; D013696:Temperature; D013997:Time Factors; D014783:Viscosity
D006454:Hemoglobins; D008024:Ligands; D011506:Proteins; D012997:Solvents; D002248:Carbon Monoxide; D005990:Glycerol