Sidedness of yeast plasma membrane vesicles and mechanisms of activation of the ATPase by detergents.
The binding of concanavalin A and of fluorescein 5'-isothiocyanate indicate similar amount of right-side-out and inside-out vesicles in plasma membrane vesicles from either glucose-starved or glucose-fermenting yeast cells. These vesicles contain low-activity and high-activity states of the ATPase, respectively. Unmasking of latent active sites can explain the limited ATPase activation (about 2-fold) produced by several detergents on both kinds of vesicles. On the other hand, lysophosphatidic acid (oleoyl) produces a 7-fold activation of the ATPase in vesicles from glucose-starved cells. This effect is accompanied by a change in Km of the enzyme and probably reflects a direct action of the detergent on the ATPase. A similar activation and Km change can be obtained by sonication of the vesicles, although in this case soybean phospholipids are required for maximal activity. Apparently the low-activity state of the yeast plasma membrane ATPase can be activated not only by glucose metabolism 'in vivo' (mechanism unknown) but also by some detergents and physical treatments 'in vitro'. Experiments with purified ATPase from glucose-starved cells also indicate that lysophosphatidic acid (oleoyl) specifically activates the enzyme. These results suggest a note of caution on considering the usual interpretation of the effects of detergents on membrane enzymes, which only take into account the unmasking of latent active sites.
Monk
B C
BC
European Molecular Biology Laboratory, Heidelberg, F.R.G.
Montesinos
C
C
Leonard
K
K
Serrano
R
R
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure; D003208:Concanavalin A / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D005285:Fermentation; D005947:Glucose / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D008563:Membrane Lipids / Q000502:physiology; D008854:Microscopy, Electron; D008970:Molecular Weight; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D014465:Ultrasonics
D003902:Detergents; D008563:Membrane Lipids; D003208:Concanavalin A; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D005947:Glucose