Direct renaturation of the dodecyl sulfate complexes of proteins with Triton X-100.
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 isoenzymes 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.
D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D008291:Malate Dehydrogenase / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011487:Protein Conformation; D011489:Protein Denaturation / Q000187:drug effects; D012967:Sodium Dodecyl Sulfate
D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D017830:Octoxynol; D007770:L-Lactate Dehydrogenase; D008291:Malate Dehydrogenase