Can sedimentation analysis contribute to the protein folding problem?

TL;DRAbstract

Protein folding is a spontaneous reaction commonly modeled by in vitro reconstitution experiments. Ultracentifugal analysis may be applied to characterize the initial, intermediate, and final states in the denaturation-renaturation cycle. With respect to intermediate states, the “molten globule state” and the “A-state” are considered to be of major importance. Using α-lactalbumin, lactate dehydrogenase, γII-crystallin and immunoglobulin as examples, compact intermediates observed at low pH may be analyzed by ultracentrifugation. Correcting for changes in the partial specific volume, and considering aggregation as a side reaction, previous conclusions regarding the increase in hydrodynamic radius of α-lactalbumin at pH 2 are questionable. In the case of lactate dehydrogenase, subunit dissociation and charge effects on the partial specific volume are superimposed in the sedimentation characteristics at low pH. s-values and reactivation kinetics prove that the enzyme in its “A-state” cons

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