FIGURE 2.

Purification of recombinant untagged RIMKLA (A) and RIMKLB (B) by gel filtration on Sepharose S-200. Mouse RIMKLA was produced in E. coli and purified by chromatography on DEAE-Sepharose and Q-Sepharose. The most active fractions of the latter column were applied on a Sepharose S-200 column. NAAG synthase activity (●) was measured in the fractions. RIMKLA was detected by Western blotting using anti-RIMKLA peptide antibodies. Mouse RIMKLB was produced in HEK cells and purified as RIMKLA before being loaded on the gel filtration column. The β-citrylglutamate synthase activity (●) of this enzyme was measured in fractions. mU, milliunits absorbance at 280 nm.