Maeda H, Steffen E, Engel J
Biophys Chem. 1978 Nov;9(1):57-64. doi: 10.1016/0301-4622(78)87015-x.
The dimerization reactions of complete Bence-Jones protein Au (VC-Au) and of its variable fragment (V-Au) were compared in 0.2 M (ionic strength) sodium phosphate buffer, pH 6.8 at 20 degrees C. The dimerization constant for VC-Au (6.6 X 10(4) M-1) was slightly smaller than a previously published value for the fragment (1.1 X 10(5) M-1). The reaction enthalpies were positive for both processes. Temperature jump experiments exhibited two kinetic phases. The relaxation time of the fast phase as well as its concentration dependence and amplitude were almost identical for VC-Au and V-Au. Only small differences were observed in the slow phase. These close similarities between the reactions of the two proteins demonstrate that dimerization occurs mainly via interactions between the variable domains and that the constant domains interfere very little. From the observation of two relaxation times it follows that the dimerization mechanism for both VC-Au and A-Au must include at least three reacting species. Mechanisms with an isomerization between monomers in two conformational states and a single dimer species are excluded by the data. Alternative mechanisms with a single monomeric species but isomerization between dimers give a rather unsatisfactory fit. A good fit can be obtained if it is assumed that both monomers and dimers can exist in two states. Rate constants of the association and dissociation steps are of the order of 10(7) M-1 s-1 and 10(2) s-1. Isomerization rate constants are in the range of 10 s-1.
