Figure 2.

Autoubiquitination of the LlANK fusion protein. Complete in vitro ubiquitination assays contained recombinant human 6× His-tagged E1 enzyme, recombinant Arabidopsis E2 enzyme GST-AtUBC8, recombinant GST-LlANK, and ubiquitin. Assay products were analyzed with western blot using either anti-ubiquitin or anti-GST antibodies. Omission of GST-AtUBC8 or GST-LlANK protein from the assay, as indicated above the lanes, resulted in a loss of protein ubiquitination. Asterisk (*), GST or GST-fusion fragments presumably due to partial proteolysis during protein purification and incubation.