| Autor: |
Ouided Friaa, Cécile Fradin, Sanjee Shivakumar |
| Rok vydání: |
2012 |
| Předmět: |
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| Zdroj: |
Biophysical Journal. 102(3) |
| ISSN: |
0006-3495 |
| DOI: |
10.1016/j.bpj.2011.11.3421 |
| Popis: |
The Bcl-2 family proteins regulate the initiation of apoptosis through protein-protein interactions that are often membrane dependant. The proapoptotic protein Bax is an attractive drug target to modulate apoptosis since it plays a vital role in mitochondria outer membrane permeabilization (MOMP), the point of no return in apoptosis. Bax monomers oligomerize to form pores in the membrane leading to the release of mitochondrial contents that triggers a caspase cascade killing the cell. The size and distribution of Bax pores on the membrane is poorly understood although much work has been done using liposome systems. To address this, we established a novel mitochondria-like planar membrane system with the potential to observe individual Bax molecules and single oligomeric pores. The system was evaluated to display the properties of a biological membrane and to allow binding by full length cBid and Bax labelled with fluorescent dyes. Protein binding was directly assessed using confocal microscopy and characterized by fluorescence correlation spectroscopy (FCS), fluorescence intensity distribution analysis (FIDA) and single particle detection. The binding properties of Bax to the planar membrane was compared to its binding properties to liposomes, which were assessed separately using fluorescence fluctuation methods. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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