Caspase-15 is autoprocessed at two sites that contain an aspartate residue in the P1' position.
Our recent characterization of porcine caspase-15 suggested that the amino terminus of the small catalytic subunit is formed by proteolytic processing between the consecutive aspartate residues D277 and D278. Since a charged residue (D278) is highly unusual in the P1' position of a caspase cleavage site, we further characterized the mechanism of caspase-15 autoproteolysis. Amino acid sequence alignments showed that D277 and D278 as well as another pair of aspartates, D270 and D271, were evolutionarily conserved among species of the mammalian clade Laurasiatheria. Site-directed mutations of these four residues and analysis of recombinant proteins showed that D270 was crucial for autoproteolysis whereas the three other aspartates were dispensable for separation of the catalytic subunits. Mutation of D270 prevented catalytic activation and abolished subsequent processing at D277. Together with previous reports, our results show that caspase-15, unlike all other caspases, efficiently cleaves sites with an aspartate in the P1' position.
Eckhart
Leopold
L
Department of Dermatology, Medical University of Vienna, A-1090 Vienna, Austria.
Kittel
Christian
C
Ballaun
Claudia
C
Tschachler
Erwin
E
D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000737:chemistry* / Q000235:genetics*; D001665:Binding Sites; D020169:Caspases / Q000737:chemistry* / Q000235:genetics*; D017124:Conserved Sequence / Q000235:genetics; D004789:Enzyme Activation; D019143:Evolution, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid
D001224:Aspartic Acid; D020169:Caspases