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Biology

Esaminando BCL-2 Funzione Famiglia con grandi vescicole unilamellari

Published: October 5, 2012 doi: 10.3791/4291

Materials

Name Company Catalog Number Comments
1,2-Dioleoyl-sn-Glycero-3-Phosph–thanolamine "PE" Avanti 850725C
L-α-Phosphatidylcholine (Egg, Chicken) "PC" Avanti 840051C
L-α-Phosphatidylinositol (Liver, Bovine) "PI" Avanti 840042C
L-α-Phosphatidylserine (Brain, Porcine) "PS" Avanti 8400320
Cardiolipin (Heart, Bovine - Sodium Salt) "CL" Avanti 840012C
Mini-extruder set Avanti 610023
PC membrane 0.2 mM Avanti 610006
Costar black 96 well plate Fisher Scientific 07-200-590
Caspase-8 cleaved human BID R&D Systems 882-B8-050
Human BCL-xL (minus C-terminus) R&D Systems 894-BX-050
BID/PUMA BH3 domain peptides Anaspec 61711/62404
Synergy H1 hybrid multi-mode microplate reader BioTek None

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References

  1. Chipuk, J. E., Moldoveanu, T., Llambi, F., Parsons, M. J., Green, D. R. The BCL-2 family reunion. Mol. Cell. 37, 299-310 (2010).
  2. Green, D. R. Apoptotic pathways: ten minutes to dead. Cell. 121, 671-674 (2005).
  3. Gavathiotis, E. BAX activation is initiated at a novel interaction site. Nature. 455, 1076-1081 (2008).
  4. Dewson, G. To trigger apoptosis, Bak exposes its BH3 domain and homodimerizes via BH3:groove interactions. Mol. Cell. 30, 369-380 (2008).
  5. Dewson, G. Bak activation for apoptosis involves oligomerization of dimers via their alpha6 helices. Mol. Cell. 36, 696-703 (2009).
  6. Gavathiotis, E., Reyna, D. E., Davis, M. L., Bird, G. H., Walensky, L. D. BH3-triggered structural reorganization drives the activation of proapoptotic BAX. Mol. Cell. 40, 481-492 (2010).
  7. Kuwana, T. BH3 domains of BH3-only proteins differentially regulate Bax-mediated mitochondrial membrane permeabilization both directly and indirectly. Mol. Cell. 17, 525-535 (2005).
  8. Chipuk, J. E. Mechanism of apoptosis induction by inhibition of the anti-apoptotic BCL-2 proteins. Proc. Natl. Acad. Sci. U.S.A. 105, 20327-20332 (2008).
  9. Letai, A. Distinct BH3 domains either sensitize or activate mitochondrial apoptosis, serving as prototype cancer therapeutics. Cancer Cell. 2, 183-192 (2002).
  10. Kuwana, T. lipids cooperate to form supramolecular openings in the outer mitochondrial membrane. Cell. 111, 331-342 (2002).
  11. Lovell, J. F. Membrane binding by tBid initiates an ordered series of events culminating in membrane permeabilization by Bax. Cell. 135, 1074-1084 (2008).
  12. Hsu, Y. T., Youle, R. J. Nonionic detergents induce dimerization among members of the Bcl-2 family. J Biol Chem. 272, 13829-13834 (1997).
  13. Suzuki, M., Youle, R. J., Tjandra, N. Structure of Bax: coregulation of dimer formation and intracellular localization. Cell. 103, 645-654 (2000).
  14. Du, H. BH3 domains other than Bim and Bid can directly activate Bax/Bak. J Biol Chem. 286, 491-501 (2011).
Esaminando BCL-2 Funzione Famiglia con grandi vescicole unilamellari
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Cite this Article

Asciolla, J. J., Renault, T. T.,More

Asciolla, J. J., Renault, T. T., Chipuk, J. E. Examining BCL-2 Family Function with Large Unilamellar Vesicles. J. Vis. Exp. (68), e4291, doi:10.3791/4291 (2012).

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