A single arginine residue is required for the interaction of the electron transferring flavoprotein (ETF) with three of its dehydrogenase partners.
The interaction of several dehydrogenases with the electron transferring flavoprotein (ETF) is a crucial step required for the successful transfer of electrons into the electron transport chain. The exact determinants regarding the interaction of ETF with its dehydrogenase partners are still unknown. Chemical modification of ETF with arginine-specific reagents resulted in the loss, to varying degrees, of activity with medium chain acyl-coenzyme A dehydrogenase (MCAD). The kinetic profiles showed the inactivations followed pseudo-first-order kinetics for all reagents used. For activity with MCAD, maximum inactivation of ETF was accomplished by 2,3-butanedione (4% residual activity after 120 min) and it was shown that modification of one arginine residue was responsible for the inactivation. Almost 100% restoration of this ETF activity was achieved upon incubation with free arginine. However, the same 2,3-butanedione modified ETF only possessed decreased activity with dimethylglycine-(DMGDH, 44%) and sarcosine- (SDH, 27%) dehydrogenases unlike the abolition with MCAD. Full protection of ETF from arginine modification by 2,3-butanedione was achieved using substrate-protected DMGDH, MCAD and SDH respectively. Cross-protection studies of ETF with the three dehydrogenases implied use of the same single arginine residue in the binding of all three dehydrogenases. These results lead us to conclude that this single arginine residue is essential in the binding of the ETF to MCAD, but only contributes partially to the binding of ETF to SDH and DMGDH and thus, the determinants of the dehydrogenase binding sites overlap but are not identical.
Parker
Antony R
AR
Department of Biochemistry, University College Dublin, Belfield, Dublin, Ireland. aparker3@jhmi.edu
D042964:Acyl-CoA Dehydrogenase / Q000737:chemistry; D000818:Animals; D001120:Arginine / Q000737:chemistry*; D001665:Binding Sites; D003931:Diacetyl / Q000737:chemistry; D050893:Dimethylglycine Dehydrogenase; D004579:Electron Transport; D044927:Electron-Transferring Flavoproteins / Q000737:chemistry* / Q000378:metabolism; D004583:Electrons; D007700:Kinetics; D010088:Oxidoreductases / Q000378:metabolism; D010089:Oxidoreductases, N-Demethylating / Q000737:chemistry; D011485:Protein Binding; D050892:Sarcosine Dehydrogenase; D013552:Swine; D013997:Time Factors
D044927:Electron-Transferring Flavoproteins; D001120:Arginine; D010088:Oxidoreductases; D042964:Acyl-CoA Dehydrogenase; D010089:Oxidoreductases, N-Demethylating; D050892:Sarcosine Dehydrogenase; D050893:Dimethylglycine Dehydrogenase; D003931:Diacetyl