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. 2014 Oct 24;289(51):35351–35362. doi: 10.1074/jbc.M114.593376

FIGURE 5.

FIGURE 5.

Lactase/phlorizin hydrolase LPH. a, schematic depicting LPH with predicted acid/base (Glu-1065, Glu-1538) and nucleophile (Glu-1273, Glu-1749) for both pockets III and IV. b, catalytic activity of LPH variants toward 4MU-, 4NP-, and 2,4DNP-β-d-Glc relative to mock (black, hatched, white columns, respectively). c, sodium azide-mediated rescue of 2,4DNP-β-d-Glc hydrolysis by mock (empty plasmid, open circles), wild-type LPH (closed circles), E1065G (gray triangles), E1273G (gray circles), E1538G (gray squares), E1749G (gray diamonds), and E1273G/E1749G (black squares). d, glucose liberation from lactose by LPH variants, relative to mock. e, overexpressed myc/His-tagged LPH variants in total COS-7 cells with excess ABP 1 or ABP 2 and subsequently purified via His-tag pulldown. Labeling with ABP 1 in the absence/presence of sodium azide (first and second gel), idem with ABP 2 (third and fourth gel), and LPH detected with α-myc (fifth gel). Data are the average of triplicates ± S.D., with one-way analysis of variance significance; *, p < 0.05; ***, p < 0.001.