Additional file 1.

Retention of proteins on the GST-HHR23A~ΔUBL column corresponds to their ability to interact with UBA domains. (A) Purified GST-HHR23A~ΔUBL protein was used to saturate glutathione-sepharose resin, which was in turn used to pack an HR10/10 column. Equal amounts of HA-tagged SUMO (HA-SUMO), polyhistidine-tagged ubiquitin (Ub-His) and FLAG-tagged UBL (UBL-FLAG) were loaded simultaneously onto the column and the column was resolved in PBS. Fractions were collected and analyzed by Western blotting using epitope-specific antibodies. (B) Purified GST was bound to glutathione-sepharose resin in excess, which was then used to pack an HR10/10 column. Equal amounts of HA-SUMO, Ub-His and UBL-FLAG were mixed, loaded onto the column and analyzed as described above.

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Goh et al. BMC Biochemistry 2008 9:4   doi:10.1186/1471-2091-9-4