Fig. 6.

ALPH proteins have mRNA decapping activity. A Recombinant ALPH proteins were produced in Arctic Express DE cells and purified via Nickel affinity chromatography. A Coomassie stained gel loaded with the purified proteins (red asterisk) is shown. Note that the Arctic Express DE cells contain chaperons (yellow asterisk). Chaperonin Cpn60 is often co-purified, in the case of TbALPH2 to a large extent. B The enzymes were tested for in vitro decapping activity in the presence of 1 mM of divalent ions, as indicated. A reaction without enzyme served as negative control. For each enzyme, an asterisk marks the conditions for best decapping activity. The gels are representative gels out of at least three replicates. An inactive mutant of Sphaeroforma arctica has no activity (last lane)