Solution structure of the antiapoptotic protein bcl-2

Petros, Andrew M.; Medek, A; Nettesheim, David G.; Kim, Daniel H.; Yoon, Ho Sup; Swift, Kerry M.; Matayoshi, Edmund D.; Oltersdorf, Tilman; Fesik, Stephen W.

doi:10.1073/pnas.041619798

Cited by 398 publications

(404 citation statements)

References 43 publications

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“…9,10,14 Similarly, numerous studies have demonstrated the interaction between CED-9 and EGL-1, and release and/or redistribution of CED-4 from the mitochondria. 3,12,[14][15][16] Recently, three-dimensional structures were determined for the Bcl-2 homology region of CED-9 alone, 16 and in complex with an EGL-1 fragment encompassing its BH3 domain, 14 confirming the structural similarity with the mammalian Bcl-2 prosurvival family members 17,18 and their mode of engagement with BH3 domains. [19][20][21] These structures of CED-9 suggest a mechanism by which CED-4 is released from CED-9.…”

Section: Introductionmentioning

confidence: 87%

CED-4 forms a 2 : 2 heterotetrameric complex with CED-9 until specifically displaced by EGL-1 or CED-13

et al. 2005

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The pathway to cell death in Caenorhabditis elegans is well established. In cells undergoing apoptosis, the Bcl-2 homology domain 3 (BH3)-only protein EGL-1 binds to CED-9 at the mitochondrial membrane to cause the release of CED-4, which oligomerises and facilitates the activation of the caspase CED-3. However, despite many studies, the biophysical features of the CED-4/CED-9 complex have not been fully characterised. Here, we report the purification of a soluble and stable 2 : 2 heterotetrameric complex formed by recombinant CED-4 and CED-9 coexpressed in bacteria. Consistent with previous studies, synthetic peptides corresponding to the BH3 domains of worm BH3-only proteins (EGL-1, CED-13) dissociate CED-4 from CED-9, but not from the gain-offunction CED-9 (G169E) mutant. Surprisingly, the ability of worm BH3 domains to dissociate CED-4 was specific since mammalian BH3-only proteins could not do so.

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Section: Introductionmentioning

confidence: 87%

CED-4 forms a 2 : 2 heterotetrameric complex with CED-9 until specifically displaced by EGL-1 or CED-13

et al. 2005

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“…The four BH domains are conserved throughout the Bcl-2 pro-survival proteins and fold into globular motifs with a hydrophobic groove on the surface, as determined by exploring the solution structures of Bcl-2, Bcl-xL and Bcl-w [310][311][312] . This groove is where the BH3 domain can bind to an amphipathic -helix of about 24 residues contained in Bax and Bak, as observed in solution structures of Bcl-x:Bak/Bax complexes 313,314 .…”

Section: The Pro-survival Proteinsmentioning

confidence: 99%

Role of the pro-survival molecule Bfl-1 in melanoma

Hind

Carter

Harris

et al. 2015

The International Journal of Biochemistry & Cell Biology

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Melanoma, the cancer derived from the melanocytes of the skin, is becoming an ever more common cancer in the ageing population of the developed world and displays an intrinsic resistance to current therapies. This chemo resistance makes metastatic melanoma an extremely difficult cancer to treat leading to a 5 year survival rate of just 20%. As such, it is important to unearth new potential drug targets to increase the prospects for melanoma patients.Bfl-1 is a pro-survival protein of the Bcl-2 family of apoptosis regulating proteins. It has been found to be over-expressed in a small subset of chemo resistant tumours, including melanoma. Accordingly, I characterised the molecule in melanoma cells and determined its role in protecting these cells from chemotherapy-induced apoptosis. I elucidated the expression profile and half-lives of the protein and mRNA across a panel of melanoma cell-lines and healthy melanocytes. I also confirmed, using pathway specific inhibitors, that Bfl-1 was transcriptionally regulated by the NFB pathway and concluded through pulsechase experiments that Bfl-1 protein was degraded, at least in part, by the proteasome. Subcellular fractionation and indirect immunofluorescence techniques elucidated that Bfl-1 was located mostly at the mitochondria in both resting and apoptotic cells, with a diffuse level present throughout the cytoplasm. As well as characterising the protein in both melanoma cells and healthy primary melanocytes, I determined its role in the resistance of these cells to apoptosis. Over-expressed Bfl-1 was found to protect melanoma cells from apoptosis caused by a range of chemotherapeutic agents in A375 melanoma cells, which naturally expressed very low levels of Bfl-1. Further, knock down of Bfl-1 using siRNA technology in melanoma cells revealed the dependence of these cells on Bfl-1 for their resistance to certain chemotherapeutic agents currently used in melanoma treatment, including the MEK inhibitor PD901. All together, this research contributes to our understanding of Bfl-1 and highlights it as a potentially important therapeutic target in metastatic melanoma.

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“…Discrete differences in the amino-acid composition among antiapoptotic grooves and BH3 ligands dictate the specificity of apoptotic-binding partners. 49,57,79,80,84 With an essential rule of engagement structurally defined, the mechanics of BCL-2 family interactions came into focus. On the proapoptotic side, NMR structures of BH3-only BID 85,86 and multidomain proapoptotic BAX 52 disclosed striking architectural similarities between the proponents and opponents of cell death (Figure 4c).…”

Section: Bcl-2 Family Form and Functionmentioning

confidence: 99%

BCL-2 in the crosshairs: tipping the balance of life and death

Walensky¹

2006

Cell Death Differ

226

175

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The discovery of B-cell lymphoma-2 (BCL-2) over 20 years ago revealed a new paradigm in cancer biology: the development and persistence of cancer can be driven by molecular roadblocks along the natural pathway to cell death. The subsequent identification of an expansive family of BCL-2 proteins provoked an intensive investigation of the interplay among these critical regulators of cell death. What emerged was a compelling tale of guardians and executioners, each participating in a molecular choreography that dictates cell fate. Ten years into the BCL-2 era, structural details defined how certain BCL-2 family proteins interact, and molecular targeting of the BCL-2 family has since become a pharmacological quest. Although many facets of BCL-2 family death signaling remain a mechanistic mystery, small molecules and peptides that effectively target BCL-2 are eliminating the roadblock to cell death, raising hopes for a medical breakthrough in cancer and other diseases of deregulated apoptosis.

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Solution structure of the antiapoptotic protein bcl-2

Cited by 398 publications

References 43 publications

CED-4 forms a 2 : 2 heterotetrameric complex with CED-9 until specifically displaced by EGL-1 or CED-13

CED-4 forms a 2 : 2 heterotetrameric complex with CED-9 until specifically displaced by EGL-1 or CED-13

Role of the pro-survival molecule Bfl-1 in melanoma

BCL-2 in the crosshairs: tipping the balance of life and death

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