Preferential cross-linking of the small subunit of the electron-transfer flavoprotein to general acyl-CoA dehydrogenase.
The interaction between pig liver mitochondrial electron-transfer flavoprotein (ETF) and general acyl-CoA dehydrogenase (GAD) was investigated by means of the heterobifunctional reagent N-succinimidyl 3-(2-pyridyldithio)propionate. Neither ETF or GAD contained reactive thiol groups. The substitution of 9.4 lysine residues/FAD group in GAD with pyridyl disulphide structures did not affect the catalytic activity of the enzyme. Thiol groups were introduced into ETF by thiolation with methyl 4-mercaptobutyrimidate. ETF containing 10.5 reactive thiol groups/FAD group showed undiminished electron-acceptor activity with respect to GAD. The reaction of thiolated ETF and GAD containing pyridyl disulphide structures resulted in a decreased staining intensity of the small subunit of ETF on SDS/polyacrylamide-gel electrophoresis. Preferential cross-linking of the smaller subunit of ETF to GAD did not take place when ETF was first treated with SDS, but was unaffected by reduction of GAD by octanoyl-CoA.
D044944:Acyl-CoA Dehydrogenases / Q000378:metabolism*; D000818:Animals; D002384:Catalysis; D003432:Cross-Linking Reagents; D044927:Electron-Transferring Flavoproteins; D004591:Electrophoresis, Polyacrylamide Gel; D005420:Flavoproteins / Q000378:metabolism*; D008239:Lysine / Q000032:analysis; D013388:Succinimides; D013552:Swine
D003432:Cross-Linking Reagents; D044927:Electron-Transferring Flavoproteins; D005420:Flavoproteins; D013388:Succinimides; C018151:N-succinimidyl 3-(2-pyridyldithio)propionate; D044944:Acyl-CoA Dehydrogenases; D008239:Lysine