The kinetics of conformational changes of alpha 2-macroglobulin determined by time resolved X-ray solution scattering

The rate of gross conformational change of alpha 2-macroglobulin (alpha 2M) during its proteinase trapping was directly determined for the first time using time-resolved X-ray solution scattering. Decrease of radius of gyration was observed under pseudo-first-order conditions with excess proteinases, which exhibited a monophasic time-course. The rate constants were 0.5 +/- 0.1 s-1 and 0.8 +/- 0.2 s-1 for the reaction with chymotrypsin and trypsin, respectively. There was no concentration dependence of the observed rate constants. Therefore, the rate-limiting step of the gross conformational change was not the bimolecular encounter reaction between alpha 2M and proteinases, which requires a new proposal of pre-trapping of proteinases before the gross conformational change.