Thermostability of respiratory terminal oxidases in the lipid environment.
The effect of the lipid environment on the thermostability of three respiratory terminal oxidases was determined. Cytochrome-c oxidase from beef heart and Bacillus stearothermophilus were used as representative proteins from mesophilic and thermophilic origin, respectively. Quinol oxidase from the archaeon Sulfolobus acidocaldarius represented the model for a extreme thermoacidophilic enzyme. All three integral membrane proteins were tested for their thermal inactivation in detergent and after reconstitution in liposomes composed of phospholipids of Escherichia coli or tetraether lipids from S. acidocaldarius. When preincubated at 0 degrees C, all three enzymes exhibited biphasic thermal inactivation curves. Data could be analysed according to a two-state model that defines two conformations of the enzyme, differing in their thermostability. Monophasic inactivation curves were observed when the enzymes were preincubated at higher temperatures prior to thermal inactivation. Lipids rendered the beef-heart cytochrome-c oxidase and S. acidocaldarius quinol oxidase more thermostable as compared to detergent solution. In contrast, the B. stearothermophilus oxidase, an intrinsically thermostable enzyme, was as thermostable in detergent as in the reconstituted state. These data suggest that the lipid environment can be an important factor in the thermostability of membrane proteins.
Elferink
M G
MG
Department of Microbiology, University of Groningen, Haren, The Netherlands.
Bosma
T
T
Lolkema
J S
JS
Gleiszner
M
M
Driessen
A J
AJ
Konings
W N
WN
D000818:Animals; D002417:Cattle; D003576:Electron Transport Complex IV / Q000737:chemistry*; D004795:Enzyme Stability; D004926:Escherichia coli / Q000737:chemistry; D001411:Geobacillus stearothermophilus / Q000737:chemistry; D008055:Lipids / Q000737:chemistry*; D009206:Myocardium / Q000737:chemistry; D010088:Oxidoreductases / Q000737:chemistry*; D016945:Sulfolobus acidocaldarius / Q000737:chemistry; D013696:Temperature
D008055:Lipids; D010088:Oxidoreductases; C020094:duroquinol oxidase; D003576:Electron Transport Complex IV