JCS030544 Supplementary Material

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• Supplemental Figure S1 (Adobe PDF) -

  Fig. S1 Identification of coronin 3 by peptide mass fingerprinting. The p53 protein was analyzed by peptide mass fingerprinting and identified as coronin 3. Underlined amino acids indicate the peptide sequences that were matched with the peptide masses detected by the TOF-MS analysis.

• Supplemental Figure S2 (Adobe PDF) -

  Fig. S2 Coronin 3 preferentially binds GDP-Rab27a. (A) COS-7 extracts expressing GFP-coronin 3 and FLAG-Rab27a mutants were immunoprecipitated with anti-FLAG antibody. The immunocomplex was analyzed by immunoblotting using anti-GFP and anti-FLAG antibodies. (B) COS-7 extracts expressing GFP-coronin 3 and FLAG-Rab3a mutants were immunoprecipitated with anti-GFP antibody. The immunocomplex was analyzed by immunoblotting using anti-GFP and anti-FLAG antibodies. (C) COS-7 extracts expressing GFP-coronin 3 and FLAG-Rab3a-T36N, -Rab5a-S34N or -Rab27a-T23N were immunoprecipitated with anti-GFP antibody. The immunocomplex was analyzed by immunoblotting using anti-GFP and anti-FLAG antibodies. (D) MIN6 extracts were immunoprecipitated with anti-Rab27a antibody. The immunocomplex was analyzed by immunoblotting using anti-Rab27a and anti-coronin 2 antibodies. (E) COS-7 extracts expressing GFP-coronin and FLAG-Rab27-T23N were immunoprecipitated with anti-GFP antibody. The immunocomplex was analyzed by immunoblotting using anti-FLAG antibody. (F) An in vitro binding assay using GST-Rab3a-T36N and various concentrations of MBP-coronin 3-ΔC. In the co-immunoprecipitation experiments, 0.2% of the input protein was co-immunoprecipitated except T36N (0.02%).

• Supplemental Figure S3 (Adobe PDF) -

  Fig. S3 Coronin 3-ΔC inhibits the uptake of anti-phogrin antibody. MIN6 cells were infected with recombinant adenoviruses bearing either LacZ (left) or coronin 3-ΔC (right) cDNA. The cells were incubated with anti-phogrin antibody, which reacts with the extracellular domain. The uptake of anti-phogrin antibody was examined by immunostaining. Scale bar, 20 µm.