title: Selective proteolysis of the protein X subunit of the bovine heart pyruvate dehydrogenase complex. Effects on dihydrolipoamide dehydrogenase (E3) affinity and enzymic properties of the complex.
abstract: Selective proteolysis of the protein X subunit of native bovine heart pyruvate dehydrogenase complex may be accomplished without loss of overall complex activity. Partial loss of function occurs if Mg2+ and thiamin pyrophosphate are not present during proteinase arg C treatment as these cofactors are necessary to prevent cleavage of the E1 alpha subunit. Specific degradation of component X leads to marked alterations in the general enzymic properties of the complex. Lipoamide dehydrogenase (E3) exhibits a decreased affinity for the core assembly and the complex is much more susceptible to inactivation at high ionic strength. The inactive form of the complex is not readily re-activated by removal of salt. It appears that intact protein X and specifically the presence of its cleaved lipoyl domain is not essential for maintenance of an enzymically active pyruvate dehydrogenase complex. However, this protein has an important structural role in promoting the correct association of E3 with the E2 core assembly, an interaction that is required for optimal catalytic efficiency of the complex.
authors.affiliation: Department of Biochemistry, University of Glasgow, Scotland, U.K.
authors.forename: J C
authors.initials: JC
authors.lastname: Neagle
authors.forename: J G
authors.initials: JG
authors.lastname: Lindsay
mesh_terms: D000818:Animals; D002417:Cattle; D008058:Dihydrolipoamide Dehydrogenase / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008274:Magnesium / Q000378:metabolism; D009206:Myocardium / Q000201:enzymology*; D009994:Osmolar Concentration; D010766:Phosphorylation; D011768:Pyruvate Dehydrogenase Complex / Q000378:metabolism*; D013379:Substrate Specificity; D013835:Thiamine Pyrophosphate / Q000378:metabolism
chemical_list: D011768:Pyruvate Dehydrogenase Complex; D008058:Dihydrolipoamide Dehydrogenase; D008274:Magnesium; D013835:Thiamine Pyrophosphate