Annexins V and XII alter the properties of planar lipid bilayers seen by conductance probes.
Annexins are proteins that bind lipids in the presence of calcium. Though multiple functions have been proposed for annexins, there is no general agreement on what annexins do or how they do it. We have used the well-studied conductance probes nonactin, alamethicin, and tetraphenylborate to investigate how annexins alter the functional properties of planar lipid bilayers. We found that annexin XII reduces the nonactin-induced conductance to approximately 30% of its original value. Both negative lipid and approximately 30 microM Ca(2+) are required for the conductance reduction. The mutant annexin XIIs, E105K and E105K/K68A, do not reduce the nonactin conductance even though both bind to the membrane just as wild-type does. Thus, subtle changes in the interaction of annexins with the membrane seem to be important. Annexin V also reduces nonactin conductance in nearly the same manner as annexin XII. Pronase in the absence of annexin had no effect on the nonactin conductance. But when added to the side of the bilayer opposite that to which annexin was added, pronase increased the nonactin-induced conductance toward its pre-annexin value. Annexins also dramatically alter the conductance induced by a radically different probe, alamethicin. When added to the same side of the bilayer as alamethicin, annexin has virtually no effect, but when added trans to the alamethicin, annexin dramatically reduces the asymmetry of the I-V curve and greatly slows the kinetics of one branch of the curve without altering those of the other. Annexin also reduces the rate at which the hydrophobic anion, tetraphenylborate, crosses the bilayer. These results suggest that annexin greatly reduces the ability of small molecules to cross the membrane without altering the surface potential and that at least some fraction of the active annexin is accessible to pronase digestion from the opposite side of the membrane.
Sokolov
Y
Y
Department of Physiology and Biophysics, University of California, Irvine, Irvine, California 92697-4560, USA.
Mailliard
W S
WS
Tranngo
N
N
Isas
M
M
Luecke
H
H
Haigler
H T
HT
Hall
J E
JE
D000408:Alamethicin / Q000494:pharmacology; D017304:Annexin A5 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D004553:Electric Conductivity; D007473:Ion Channels / Q000502:physiology; D007536:Isomerism; D008051:Lipid Bilayers / Q000378:metabolism*; D018942:Macrolides / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D016297:Mutagenesis, Site-Directed / Q000502:physiology; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D012965:Sodium Chloride / Q000494:pharmacology; D013775:Tetraphenylborate / Q000493:pharmacokinetics; D014475:Uncoupling Agents / Q000494:pharmacology
D017304:Annexin A5; D007473:Ion Channels; D008051:Lipid Bilayers; D018942:Macrolides; D014475:Uncoupling Agents; D000408:Alamethicin; D013775:Tetraphenylborate; D012965:Sodium Chloride; D002118:Calcium; C050476:nonactin