[Interaction of NADH-dehydrogenase from M. lysodeikticus membranes with lipids in a reconstituted system].
The NADH-dehydrogenase isolated from the M. lysodeikticus membranes was reconstituted into liposomes from the lipids obtained from the same membranes. The presence and degree of the reconstitution were investigated by two-dimensional immunoelectrophoresis and photoreactive hydrophobic label. The quenching of protein fluorescence by the aqueous quencher J- was practically the same for the enzyme in the reconstituted system and in the detergent solution, whereas the quencher interacting with the membrane--cetylpyridinium chloride--was effective in the first case and not effective in the second one. Evidence for the energy transfer from protein chromophores of NADH dehydrogenase in the proteoliposomes (lambda excit = 286 nm) to the hydrophobic fluorescent probe pyrene was obtained. It was found that about 30% of the chromophores in the enzyme molecule are involved in this process. The hydrophobic spin probe, whose paramagnetic fragment is located on the surface and not inside the hydrophobic phase of the membrane, can act as electron acceptor during NADH oxidation in the reconstituted system. The data obtained are suggestive of the exposure of the bulk of the enzyme molecule to the environment and of interaction of the smaller part of the molecule with the lipid phase. The active center is located on the part of the enzyme molecule which is exposed to water. It is assumed that the NADH-dehydrogenase molecule is exposed to water. It is assumed that the NADH-dehydrogenase molecule is involved in heat diffusion which facilitates the active center interaction with the membrane surface.
D002462:Cell Membrane / Q000201:enzymology; D003579:Cytochrome Reductases / Q000378:metabolism*; D007122:Immunoelectrophoresis; D007700:Kinetics; D008081:Liposomes; D046911:Macromolecular Substances; D008563:Membrane Lipids / Q000502:physiology*; D008837:Micrococcus / Q000201:enzymology*; D009245:NADH Dehydrogenase / Q000378:metabolism*; D010743:Phospholipids / Q000502:physiology
D008081:Liposomes; D046911:Macromolecular Substances; D008563:Membrane Lipids; D010743:Phospholipids; D003579:Cytochrome Reductases; D009245:NADH Dehydrogenase
Issledovanie vzaimodeĭstviia NADH-degidrogenazy iz membran M. lysodeikticus s lipidami v rekonstruirovannoĭ sisteme.