Direct renaturation of the dodecyl sulfate complexes of proteins with triton X-100

Abstract

A simple procedure is described for renaturing dodecyl sulfate-unfolded enzymes. The method involves the direct addition of a large molar excess of the non-ionic detergent Triton X-100 to protein-dodecyl sulfate complexes either in solution or as a band on a polyacrylamide gel. The cytoplasmic enzymes lactate dehydrogenase and malate dehydrogenase have been renatured by this protocol. On the other hand, no recovery of activity was found with the mitochondrial isoenzyme of malate dehydrogenase or the mitochondrial enzymes glutamate dehydrogenase or fumarase. Possible implications of the differences in the ability of cytosolic and mitochondrial enzymes to renature under these conditions are discussed in terms of their biosynthesis.