Phospholipid specificity of bovine heart bc1 complex.
Bovine heart bc1 complex was reversibly inactivated by a new simple and effective chromatographic delipidation method. Upon phospholipid replenishment, catalytic activity increased from values near zero to values 2-6-times higher than those of the original preparation. Compared to original preparations maximally activated by additional phospholipid, the degree of reactivation was up to 100%. By this delipidation method, the 6.4-kDa protein subunit was removed with the phospholipid. The loss of this protein neither diminished electron transport activity nor abolished proton translocation. Two requirements were necessary to obtain quantitative data: (a) only bc1 complexes, homogeneously dissolved before and after relipidation had to be used and (b) the phospholipid bound to the complex had to be determined. The correlation of catalytic activity to bound phospholipid was studied in the range of low phospholipid/protein ratios, which had previously been insufficiently resolved. Catalytic activity increased linearly with added phospholipid up to a molar ratio of 80-100 lipid molecules/dimeric complex. This corresponds to the number of phospholipid molecules that complete a single bilayer annulus. The activating effect of phospholipid is not merely due to a hydrophobic phase effect, since it strongly depends on the nature of the polar head group of the added phospholipid. Of the three major phospholipids bound to the bc1 complex, only phosphatidylethanolamine and phosphatidylcholine activated when added as sole phospholipid. Tightly bound diphosphatidylglycerol was needed for preservation of the native complex structure.
Schägger
H
H
Gustav-Embden Zentrum der Biologischen Chemie, Johann Wolfgang Goethe-Universität, Frankfurt, Federal Republic of Germany.
Hagen
T
T
Roth
B
B
Brandt
U
U
Link
T A
TA
von Jagow
G
G
D000818:Animals; D001665:Binding Sites; D002384:Catalysis; D002417:Cattle; D002499:Centrifugation, Density Gradient; D004579:Electron Transport; D014450:Electron Transport Complex III / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D004789:Enzyme Activation / Q000187:drug effects; D004793:Enzyme Reactivators / Q000032:analysis; D009206:Myocardium / Q000201:enzymology*; D017830:Octoxynol; D010715:Phosphatidylglycerols / Q000502:physiology; D010741:Phospholipases A / Q000494:pharmacology; D010743:Phospholipids / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology; D010738:Type C Phospholipases / Q000494:pharmacology
D004793:Enzyme Reactivators; D010715:Phosphatidylglycerols; D010743:Phospholipids; D011092:Polyethylene Glycols; D017830:Octoxynol; D010741:Phospholipases A; D010738:Type C Phospholipases; D014450:Electron Transport Complex III