Comparative Proteomic Analysis Identifies a Role for SUMO in Protein Quality Control

Michael H. Tatham, Ivan Matic, Matthias Mann, Ronald T. Hay*

*To whom correspondence should be addressed. E-mail: r.t.hay{at}dundee.ac.uk

This PDF file includes:

• Materials and Methods
• Fig. S1. TAP–SUMO-2 conjugation response to MG132 treatment under SILAC conditions.
• Fig. S2. Filtering TAP–SUMO-2 protein identifications and normalizing with SILAC ratio data from crude cell lysates.
• Fig. S3. Known SUMO substrates are enriched in the list of putative SUMO-2 substrates in comparison with those proteins rejected from the analysis.
• Fig. S4. "Putative substrates" run in SDS-PAGE gels at molecular weights consistent with posttranslational modification, whereas "internal rejects" do not.
• Fig. S5. Changes in protein ratios during MG132 treatment are due to altered SUMOylation and not changes in overall protein concentrations.
• Fig. S6. Only a small proportion of the total cellular pool of most substrates is modified by SUMO-2 and responds to MG132 with altered SUMOylation.
• Fig. S7. Lactacystin and MG132 trigger similar changes in SUMO-2 conjugation in HeLa cells.
• Fig. S8. Comparisons of the functions of SUMO-2 substrates identified here and in a previous study.
• Fig. S9. Canavanine induces SUMO-2/3 conjugation and turnover.
• Fig. S10. Comparison between the effects of heat shock and MG132 on proteins involved in SUMO and ubiquitin metabolism purified by TAP–SUMO-2 from HeLa cells.
• Fig. S11. Slice-by-slice analysis of data shown in Fig. 6A.
• Fig. S12. Slice-by-slice analysis of data shown in Fig. 6, B and C.
• Fig. S13. Comparison of the quantitative data between proteins common to this study and that of Schimmel et al.
• Fig. S14. A working model for the response of SUMO and ubiquitin to proteasome inhibition.
• Fig. S15. The SUMO conjugation signal in response to MG132 transfers from the cytoplasm to the nucleus.
• Table S1. Frequency analysis of SUMO conjugation consensus sequences from proteins identified in this study compared with those of previous studies.
• Descriptions of Files 1 to 6
• References
  

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© 2011 American Association for the Advancement of Science