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Gene Synthesis> | ... actin, Arp4, Arp5, Arp8, Ies2, Ies6, Tip49a and Tip49b, were synthesized with codon bias for expression in insect cells (Genscript). A further truncated form of Ino80 comprising residues 487–1556 was cloned with an N-terminal 8-histidine and C-terminal twin Strep-tag® II. ... | Get A Quote |
We have purified a minimal core human Ino80 complex from recombinant protein expressed in insect cells. The complex comprises one subunit each of an N-terminally truncated Ino80, actin, Arp4, Arp5, Arp8, Ies2 and Ies6, together with a single heterohexamer of the Tip49a and Tip49b proteins. This core complex has nucleosome sliding activity that is similar to that of endogenous human and yeast Ino80 complexes and is also inhibited by inositol hexaphosphate (IP6). We show that IP6 is a non-competitive inhibitor that acts by blocking the stimulatory effect of nucleosomes on the ATPase activity. The IP6 binding site is located within the C-terminal region of the Ino80 subunit. We have also prepared complexes lacking... More