Supplemental Material

This article contains the following supporting material:

• Supplemental Figure 1 - Current model of the Toxoplasma motile apparatus. PM, plasma membrane; IMC, inner membrane complex.
• Supplemental Figure 2 - TgGAP45-YFP does not associate with other members of the glideosome complex. Toxoplasma tachyzoites expressing TgGAP45-YFP were labeled for 20 hours with ³⁵S methionine/cysteine and, after lysis in TX100/TBS, subjected to immunoprecipitation with antisera to TgGAP45 or YFP. Asterisks indicate degradation products of TgGAP45 and TgGAP45-YFP.
• Supplemental Table - Summary of the various crosslinking and immunoprecipitation conditions tested in this study. As indicated in the text, glideosome solubilization was only found to be temperature dependent using the detergent TX100 in 150 mM NaCl, 5 mM EDTA, 50 mM Na₃PO₄. Crosslinking experiments were performed using a variety of water insoluble (DSP, DSS, BMH) and water soluble (BS3) crosslinkers. Reactions were carried out for various times at room temperature and ice in phosphate-buffered saline (PBS) and quenched by the addition of 1M Tris or glycine, pH 7.5 to a final concentration of 50mM.