title: Studies of the structure-function relationships of Neurospora pyruvate kinase: denaturation by urea and the effect of ligands on the refolding and renaturation process.
abstract: 1. The renaturation of Neurospora pyruvate kinase following denaturation with urea was investigated. 2. The substrates, phosphoenol pyruvate and adenosine diphosphate, were observed to stabilize the enzyme against urea-induced structural disorganization. 3. It was demonstrated that for refolding/reactivation of the denatured protein the allosteric activator, fructose-1,6-diphosphate and a sulphydryl protectant are required. 4. The enzyme recovered following renaturation showed complete immunological identity with the native enzyme in Ouchterlony double diffusion tests.
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D004789:Enzyme Activation; D008024:Ligands; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology*; D010728:Phosphoenolpyruvate / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects; D011770:Pyruvate Kinase / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014508:Urea
chemical_list: D008024:Ligands; D000244:Adenosine Diphosphate; D010728:Phosphoenolpyruvate; D014508:Urea; D011770:Pyruvate Kinase