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

Abstract

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.

Statistics

050100'04'05'06'07'08'09'10'11'12'13'14'15'16'17
Citations per Year

81 Citations

Semantic Scholar estimates that this publication has 81 citations based on the available data.

See our FAQ for additional information.

Cite this paper

@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} }