Fig. 3

In silico analysis of tubulin isoforms and conserved Lys40 in alpha tubulin. a Clustal omega analysis showing Lys 40 of Alpha tubulin whereas, Beta and Gamma tubulins have serine and alanine respectively. b Lys40 position was found to be conserved in eukaryotes. c 3D crystal structures (generated by homology modelling) of Alpha, Beta and Gamma tubulins, used for studying protein protein interactions with HDAC8. d Protein protein docking studies of HDAC8 with Alpha, Beta and Gamma tubulins. e Fluorescence spectroscopy results of (a) GST-HDAC8 (30 μg) + ac-alpha tubulin peptide (500 nM) incubation from 5 to 30 min, decreases intrinsic fluorescence. (b) GST-HDAC8 (30 μg) + Unac-alpha tubulin peptide (500 nM) incubation from 5 to 30 min, no change in intrinsic fluorescence. f CD results for change in secondary structure for GST-HDAC8 alone, GST-HDAC8 (2 μM) + ac-alpha tubulin peptide (500 nM), and GST-HDAC8 (2 μM) + Unac-alpha tubulin peptide (500 nM), significant change in the secondary structure of GST-HDAC8 upon incubating with ac-alpha tubulin clearly signifies the more binding specificity for ac-alpha tubulin but not with unacetylated form