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. 2009 Oct 29;284(52):36377–36386. doi: 10.1074/jbc.M109.057257

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© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 8. — Effects of DdCAD-1 conformational changes on the import process. A, His₆-DdCAD-1 was denatured by boiling for 10 min followed by quick cooling on ice. The denatured form of His₆-DdCAD-1 and the penta-mutant (N58A, E59A, D61A, D201A, and N202A) protein were subjected to the in vitro import assay. B, effects of the tryptophan mutations, W37A and W55A, on DdCAD-1 import into contractile vacuoles were examined. Protein blots were probed with anti-DdCAD-1 antibody. C, import of DdCAD-1 dimers into contractile vacuoles. His₆-DdCAD-1 was cross-linked using disuccinimidyl suberate (DSS) and then subjected to the import assay. Protein blots were probed with anti-DdCAD-1 antibody.