Structure of the cytochrome c oxidase complex: labeling by hydrophilic and hydrophobic protein modifying reagents.
Beef heart cytochrome c oxidase has been reacted with [35S]diazobenzenesulfonate ([35S]DABS), [35S]-N-(4-azido-2-nitrophenyl)-2-aminoethylsulfonate ([35S]NAP-taurine), and two different radioactive arylazidophospholipids. The labeling of the seven different subunits of the enzyme with these protein modifying reagents has been examined. DABS, a water-soluble, lipid-insoluble reagent, reacted with subunits II, III, IV, V, and VII but labeled I or VI only poorly. The arylazidophospholipids, probes for the bilayer-intercalated portion of cytochrome c oxidase, labeled I, III, and VII heavily and II and IV lightly but did not react with V or VI. NAP-taurine labeled all of the subunits of cytochrome c oxidase. Evidence is presented that this latter reagent reacts with the enzyme from outside the bilayer, and the pattern of labeling with the different hydrophilic and hydrophobic labeling reagents is used to derive a model for the arrangement of subunits in cytochrome c oxidase.
D000818:Animals; D001557:Benzenesulfonates; D001665:Binding Sites; D002417:Cattle; D003979:Diazonium Compounds; D003576:Electron Transport Complex IV / Q000378:metabolism; D007202:Indicators and Reagents; D007553:Isotope Labeling; D009206:Myocardium / Q000201:enzymology; D010743:Phospholipids; D011485:Protein Binding; D013425:Sulfanilic Acids / Q000031:analogs & derivatives; D013462:Sulfur Radioisotopes; D013654:Taurine / Q000031:analogs & derivatives*
D001557:Benzenesulfonates; D003979:Diazonium Compounds; D007202:Indicators and Reagents; D010743:Phospholipids; D013425:Sulfanilic Acids; D013462:Sulfur Radioisotopes; D013654:Taurine; C010861:NAP taurine; D003576:Electron Transport Complex IV; C000469:diazobenzenesulfonic acid