Evolutionary Relationships and Protein Domain Architecture in an Expanded Calpain Superfamily in Kinetoplastid Parasites
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Employing whole-genome analysis we have characterized a large family of genes coding for calpain-related proteins in three kinetoplastid parasites. We have defined a total of 18 calpain-like sequences in Trypanosoma brucei, 27 in Leishmania major, and 24 in Trypanosoma cruzi. Sequence characterization revealed a well-conserved protease domain in most proteins, although residues critical for catalytic activity were frequently altered. Many of the proteins contain a novel N-terminal sequence motif unique to kinetoplastids. Furthermore, 24 of the sequences contain N-terminal fatty acid acylation motifs indicating association of these proteins with intracellular membranes. This extended family of proteins also includes a group of sequences that completely lack a protease domain but is specifically related to other kinetoplastid calpain-related proteins by a highly conserved N-terminal domain and by genomic organization. All sequences lack the C-terminal calmodulin-related calcium-binding domain typical of most mammalian calpains. Our analysis emphasizes the highly modular structure of calpains and calpain-like proteins, suggesting that they are involved in diverse cellular functions. The discovery of this surprisingly large family of calpain-like proteins in lower eukaryotes that combines novel and conserved sequence modules contributes to our understanding of the evolution of this abundant protein family.
KeywordsCalpain Trypanosoma Leishmania Parasites Genome Multigene family Protein domains Microtubules Cytoskeleton Acylation
We thank David Lunt (Hull University) for constructive discussions concerning phylogenetic analysis and Robin Allaby (Astra Zeneca) for help during the initial phase of this project. This work was funded by a Wellcome Trust Programme Grant to K.G. and a University of Hull Start-Up Grant to K.E. Kinetoplastid genomic data were accessed via http://www.genedb.org/. Most of the genomic data were provided by the Wellcome Trust Sanger Institute and The Institute for Genomic Research (TIGR), which are supported by the Wellcome Trust and the National Institutes of Health, USA (Grant AI043062), respectively.
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