Figure 4: Catalytic activity of D14 in the interaction with SLR1.
From: Molecular mechanism of strigolactone perception by DWARF14
(a) Enzymatic degradation of (±)-GR24 by D14. (±)-GR24 (10 μM) was incubated with or without purified D14 for 1 h at 37 °C. The reaction solutions were extracted with ethyl acetate, and the organic layer was subjected to HPLC analyses using a Chiralpak IA-3 column. Peaks A and B were confirmed to represent (−)-ent-GR24 (5) and (+)-GR24 (2), respectively, as described in the Methods section. The experiments were repeated three times with similar results. (b) Growth of AH109 transformants with pGAD-SLR1 and the wild-type or mutated D14 cDNA on pGBK-T7 on an SD-His, Ade plate with or without 10 μM (−)-ent-2′-epi-GR7 (3). (c) Comparison of the specific binding of T2-GR7 (9) to wild-type D14 and to H297A mutant D14 (mean±s.d., n=3).
