title: Binding of the human "electron transferring flavoprotein" (ETF) to the medium chain acyl-CoA dehydrogenase (MCAD) involves an arginine and histidine residue.
abstract: The interaction between the "electron transferring flavoprotein" (ETF) and medium chain acyl-CoA dehydrogenase (MCAD) enables successful flavin to flavin electron transfer, crucial for the beta-oxidation of fatty acids. The exact biochemical determinants for ETF binding to MCAD are unknown. Here we show that binding of human ETF, to MCAD, was inhibited by 2,3-butanedione and diethylpyrocarbonate (DEPC) and reversed by incubation with free arginine and hydroxylamine respectively. Spectral analyses of native ETF vs modified ETF suggested that flavin binding was not affected and that the loss of ETF activity with MCAD involved modification of one ETF arginine residue and one ETF histidine residue respectively. MCAD and octanoyl-CoA protected ETF against inactivation by both 2,3-butanedione and DEPC indicating that the arginine and histidine residues are present in or around the MCAD binding site. Comparison of exposed arginine and histidine residues among different ETF species, however, indicates that arginine residues are highly conserved but that histidine residues are not. These results lead us to conclude that this single arginine residue is essential for the binding of ETF to MCAD, but that the single histidine residue, although involved, is not.
authors.affiliation: Department of Biochemistry, University College Dublin, Belfield, Dublin 4, Ireland. aparker3@jhmi.edu
authors.forename: Antony R
authors.initials: AR
authors.lastname: Parker
mesh_terms: D042964:Acyl-CoA Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D001120:Arginine / Q000378:metabolism*; D001665:Binding Sites; D003931:Diacetyl / Q000494:pharmacology; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D044927:Electron-Transferring Flavoproteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D006639:Histidine / Q000031:analogs & derivatives* / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D011485:Protein Binding / Q000187:drug effects; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D013053:Spectrophotometry
chemical_list: D044927:Electron-Transferring Flavoproteins; C072750:N-carbethoxyhistidine; D006639:Histidine; D001120:Arginine; D042964:Acyl-CoA Dehydrogenase; D003931:Diacetyl; D004047:Diethyl Pyrocarbonate