Volume 8 Issue 8 August 2007
A family of aspartic proteases and a novel, dynamic, and cell-cycle dependent protease localization in the secretory pathway of Toxoplasma gondii
Michael Shea, Ursula Jäkle, Qing Liu, Colin Berry, Keith A. Joiner, and Dominique Soldati

Supplemental Figure 1. Alignment of TgASPs and plasmepsins Red bar indicates catalytic site, blue bar shows flap, brown bar shows polyproline loop, yellow bar shows TgASP1-specific insert. The YASLL sequence of TgASP1 is boxed in red.

Figure 1 (.pdf)

Supplemental Figure 2. Alignment of all apicomplexan aspartic proteases used for the phylogenetic analysis. Regions used in phylogenetic analysis are: residues 190-222, 267-280, 300-320, 400-480, 508-527, 537-577, 589-599, 606-620, 627-633, 650-678, 720-745, 806-818, 826-835, 936-978, 996-1032, 1047-1057.

Figure 2 (.pdf)

Supplemental Figure 3. Apicomplexan ASPs fall into five phylogenetic groups with little overlap with ASPs of related species. A phylogenetic tree of apicomplexan ASPs, as well as those of the related non-apicomplexans Thalassiosira pseudonana and Phytophthora ramorum and human ASPs was generated using PAUP*4.0b10. Apicomplexan ASPs cluster primarily in the same five groups identified in Figure 2.

Figure 3 (.pdf)

Supplemental Figure 4. Epitope-tagged TgASP1 shows a similar localization to wild-type TgASP1. S4A. IFA was performed on strain RHmycASP1 using α-myc antibodies, and on strain RH&dhelta;hxgprt using α-peptide2 antibody. S4B. IFA was performed on strain RHmycASP1Ty1 using α-Ty1 and α-peptide2 antibodies; merge is shown on the right hand panel.

Figure 4 (.pdf)

Supplemental Table 1. Transcription of the different TgASP genes is developmentally regulated. The table lists the number of expressed sequence tags (ESTs) present for different life-cycle stages for the different TgASPs (taken from ToxoDB and ApiDots databases). Detection or not of transcripts in sporozoites by SAGE analysis is indicated. The right hand column indicates whether the cDNA could be amplified from tachyzoites total RNAs.

Supplemental Table 2. Primers and restriction sites used in generating DNA constructs.