Figure 2.
Domain engineering and reconstitution strategy for UGT74B1/UGT74C1.A, 3D models of UGT74B1 and UGT74C1. Domains are colored as in panel A (green: NB1; blue: CB1; pink: NC1; yellow: CC1), the linker is depicted as a red loop, the red arrow marks the site of separation as in panel A. B, UGT74B1 and UGT74C1 3D domain organization. The red arrow marks the site of separation that was used in the domain cloning strategy. C, SDS PAGE analysis eluted fractions after Ni-NTA purification. Expected domain sizes are respectively 30 kDa (NB1), 31 kDa (NC1), 23 kDa (CB1), and 23 kDa (CC1). This gel was also used for peptide mass fingerprinting for identification of the chimer domains (see Figs. S3 and S4).