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Figure 2


Fig. 2. Amino acid sequence of {alpha}-tubulin and western blot analysis of {alpha}-tubulin from S100 cell extracts. (A) The amino acid sequence of human {alpha}-tubulin (TUBA3). Solid boxes indicated the tryptic peptides that were identified by mass spectrometry analysis. Arrows indicate putative grB cleavage sites. (B) HeLa S100 (lanes 1-6) or Jurkat S100 (lanes 7-12) were treated with the indicated amounts of purified grB (in a 10 µl reaction volume), in the presence or absence of 100 µM zVAD-fmk. Protein products were separated by 12% SDS-PAGE and transferred to nitrocellulose. The migration of {alpha}-tubulin (upper) and caspase 3 (lower) were assessed by western blot analysis. Full-length {alpha}-tubulin is indicated by *, and cleaved {alpha}-tubulin by **. GrB-dependent processing of pro-caspase 3 is indicated by p20, and caspase-dependent processing is indicated by p19.





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