G. Basanez et al., Bax, but not Bcl-x(L), decreases the lifetime of planar phospholipid bilayer membranes at subnanomolar concentrations, P NAS US, 96(10), 1999, pp. 5492-5497
Citations number
35
Categorie Soggetti
Multidisciplinary
Journal title
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Release of proteins through the outer mitochondrial membrane can be a criti
cal step in apoptosis, and the localization of apoptosis-regulating Bcl-2 f
amily members there suggests they control this process. We used planar phos
pholipid membranes to test the effect of full-length Bax and Bcl-x(L) synth
esized in vitro and native Bax purified from bovine thymocytes. Instead of
forming pores with reproducible conductance levels expected for ionic chann
els, Bax, but not Bcl-xL, created arbitrary and continuously variable chang
es in membrane permeability and decreased the stability of the membrane, re
gardless of whether the source of the protein was synthetic or native, This
breakdown of the membrane permeability barrier and destabilization of the
bilayer was quantified by using membrane lifetime measurements. Bax decreas
ed membrane lifetime in a voltage- and concentration-dependent manner. Bcl-
x(L) did not protect against Bax-induced membrane destabilization, supporti
ng the idea that these two proteins function independently. Corresponding t
o a physical theory for lipidic pore formation, Bax potently diminished the
linear tension of the membrane (i.e., the energy required to form the edge
of a new pore). We suggest that Bax acts directly by destabilizing the lip
id bilayer structure of the outer mitochondrial membrane, promoting the for
mation of a pore-the apoptotic pore-large enough to allow mitochondrial pro
teins such as cytochrome c to be released into the cytosol, Bax could then
enter and permeabilize the inner mitochondrial membrane through the same ho
le.