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Mol Biol Cell. 2015 Jan 15;26(2):339-49. doi: 10.1091/mbc.E13-11-0638. Epub 2014 Nov 19.

Visual and functional demonstration of growing Bax-induced pores in mitochondrial outer membranes.

Author information

  • 1La Jolla Institute, La Jolla, CA 92037.
  • 2Department of Pathology, University of Iowa, Iowa City, IA 52242.
  • 3La Jolla Institute, La Jolla, CA 92037 tomomi@lji.org.

Abstract

Bax induces mitochondrial outer membrane permeabilization (MOMP), a critical step in apoptosis in which proteins are released into the cytoplasm. To resolve aspects of the mechanism, we used cryo-electron microscopy (cryo-EM) to visualize Bax-induced pores in purified mitochondrial outer membranes (MOMs). We observed solitary pores that exhibited negative curvature at their edges. Over time, the pores grew to ∼ 100-160 nm in diameter after 60-90 min, with some pores measuring more than 300 nm. We confirmed these results using flow cytometry, which we used to monitor the release of fluorescent dextrans from isolated MOM vesicles. The dextran molecules were released gradually, in a manner constrained by pore size. However, the release rates were consistent over a range of dextran sizes (10-500 kDa). We concluded that the pores were not static but widened dramatically to release molecules of different sizes. Taken together, the data from cryo-EM and flow cytometry argue that Bax promotes MOMP by inducing the formation of large, growing pores through a mechanism involving membrane-curvature stress.

PMID:
25411335
PMCID:
PMC4294680
DOI:
10.1091/mbc.E13-11-0638
[PubMed - indexed for MEDLINE]
Free PMC Article
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