Nonideality and the nucleation of sickle hemoglobin.
The homogeneous and heterogeneous nucleation kinetics of sickle hemoglobin (HbS) have been studied for various degrees of solution crowding by substitution of cross-linked hemoglobin A, amounting to 50% of the total hemoglobin. By cross-linking hemoglobin A, hybrid formation between hemoglobin A and hemoglobin S was prevented, thus simplifying the analysis of the results. Polymerization was induced by laser photolysis, and homogeneous nucleation kinetics were determined by observation of the stochastic behavior of the onset of light scattering. Heterogeneous nucleation was determined by observing the exponential growth of the progress curves, monitored by light scattering. At concentrations between 4 and 5 mM tetramer (i.e., approximately 30 g/dl), the substitution of 50% HbA for HbS slows the reaction by a factor of 10(3) to 10(4). Using scaled particle theory to account for the crowding of HbA, the observed decrease in the homogeneous nucleation rate was accurately predicted, with no variation of parameters required. Heterogeneous nucleation, on the other hand, is not well described in the present formulation, and the theory for this process appears to require modification of the way in which nonideality is introduced. Nonetheless, the accuracy of the homogeneous nucleation description suggests that such an approach may be useful for other assembly processes that occur in a crowded intracellular milieu.
Ivanova
M
M
Department of Physics, Drexel University, Philadelphia, Pennsylvania 19104, USA.
Jasuja
R
R
Kwong
S
S
Briehl
R W
RW
Ferrone
F A
FA
D003432:Cross-Linking Reagents; D006441:Hemoglobin A / Q000737:chemistry*; D006451:Hemoglobin, Sickle / Q000737:chemistry*; D006801:Humans; D007700:Kinetics; D010782:Photolysis; D055503:Protein Multimerization; D012996:Solutions
D003432:Cross-Linking Reagents; D006451:Hemoglobin, Sickle; D012996:Solutions; D006441:Hemoglobin A