Analysis of the minimal specificity of CED-3 using a yeast transcriptional reporter system.
Determination of the substrate specificity of site-specific proteases helps define their physiological roles. We developed a yeast-based system for defining the minimal substrate specificity of site-specific proteases, within the context of a protein. Using this system, we characterized the P4-P1 substrate specificity of the nematode apoptotic caspase CED-3. Apart from an absolute requirement for aspartate at the P1 position, CED-3 is a relatively promiscuous caspase capable of cleaving substrates bearing many amino acids at P4-P2 sites.
Westein
Sarah J
SJ
Department of Biochemistry, La Trobe University, Bundoora, Victoria, Australia.
Scott
Fiona L
FL
Hawkins
Christine J
CJ
D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D001665:Binding Sites; D029742:Caenorhabditis elegans Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D020169:Caspases / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D008954:Models, Biological; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000378:metabolism; D020539:Sequence Analysis, Protein; D013379:Substrate Specificity; D014158:Transcription, Genetic; D014162:Transfection; D015003:Yeasts / Q000235:genetics*
D029742:Caenorhabditis elegans Proteins; D011994:Recombinant Proteins; D001224:Aspartic Acid; D020169:Caspases; C089223:ced-3 protein, C elegans