Subcellular localization, oligomerization, and ATP-binding of Caenorhabditis elegans CED-4.


Caspase family cell death proteases are activated during apoptosis through the oligomerization of caspase-binding "adapter" proteins. In the nematode Caenorhabditis elegans one adapter protein, CED-4, exists. Here we report an analysis of CED-4 protein expressed in insect Sf9 cells by infection with recombinant baculovirus. During expression, CED-4 assumed a perinuclear spherical or reticular localization where it was partly resistant to extraction with nonionic detergents. Both purified FLAG-CED-4 and GST-FLAG-CED-4 proteins were present in solution as large complexes. FLAG-CED-4 complexes were estimated by gel filtration to have a molecular weight of approximately 500 kDa to >1.2 MDa, while GST-FLAG-CED-4 complexes appeared somewhat smaller. Unlike its mammalian homologue Apaf-1, CED-4 exhibited a marked preference for ATP over dATP in filter binding studies and in competition experiments. ATP hydrolysis was required neither for complex stability nor for binding of CED-3. These features are likely to be relevant for CED-4's function as a caspase adapter.


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@article{Seiffert2002SubcellularLO, title={Subcellular localization, oligomerization, and ATP-binding of Caenorhabditis elegans CED-4.}, author={Barbara M. Seiffert and Juliane Vier and Georg H{\"a}cker}, journal={Biochemical and biophysical research communications}, year={2002}, volume={290 1}, pages={359-65} }