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. 2006 Feb;140(2):704–715. doi: 10.1104/pp.105.071514

Figure 3.

Figure 3.

Affinity purification and substrate specificity analysis of the E. coli-expressed His-tagged THT protein. A, Expression of the pepper THT gene (AY819700) in E. coli. Protein samples were separated by SDS-PAGE and stained using Coomassie Blue. Lane M, Molecular standard; lane 1, total proteins in a 10-μL aliquot of bacterial cells grown without IPTG; lane 2, total proteins in a 10-μL aliquot of bacterial cells after IPTG treatment; lane 3, 20 μg of soluble proteins; lane 4, the THT protein (10 μg) purified by affinity (Ni-NTA) chromatography. B, Substrate specificity of the purified recombinant THT protein. Footnote a, Tyramine (1 mm) was used as the acyl acceptor. Footnote b, Feruloyl-CoA (250 μm) was used as the acyl donor. nd, Not detected.