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. 2009 Mar 6;37(8):2596–2606. doi: 10.1093/nar/gkp115

Figure 2.

Figure 2.

PfHP1 binds to histone H3 peptide methylated at lysine 9. (A) Total proteins from DH5α strain expressing the CD of PfHP1 before (NI) and after IPTG induction (I), as well as the purified GST-PfHP1-CD fusion protein (P), were separated by SDS–PAGE and stained with Coomassie blue (left). An anti-GST antibody was used to detect the presence of the fusion protein (right). (B) Purified protein GST-PfHP1-CD was assayed for binding to histone H3 peptides. 200 ng of GST-CD protein (GST-PfHP1-CD) were dotted to a nitrocellulose membrane and incubated with 2 µg of the following biotinylated peptides: H3K9me3, H3K9me2, H3K9ac, H3K4me3, H3K4me2 and unmodified H3. (C) Two hundred nanograms of GST-PfHP1-CD fusion protein were dotted to nitrocellulose and incubated with decreasing amounts of H3K9me3 and H3K9me2 biotinylated peptides (left panels). A representative result of two independent experiments is shown. The right panel shows a quantitative presentation of the GST-PfHP1-CD affinity to the peptides.