Extended Data Fig. 1: Biochemical analysis of the BIC2-CRY2 complex.
From: Structural insights into BIC-mediated inactivation of Arabidopsis cryptochrome 2
a, The interactions between two BICs (BIC1 and BIC2) and two CRYs (CRY1 and CRY2) in Arabidopsis thaliana were assessed by pull-down assays. b, Characterization of the purified full-length BIC2-CRY2 complex. Upper panel is the gel filtration chromatogram for BIC2-CRY2 purified using a SuperdexTM 200 increase 10/300 GL column. Elution volume of BIC2-CRY2 = 13.7 ml. The calibration standard for gel filtration chromatography is a mixture of thyroglobulin, γ-globulin, and ovalbumin proteins with approximate molecular weights of 669 kDa (10.3 ml), 158 kDa (13.0 ml), and 44 kDa (15.7 ml), respectively. The elution volume of BIC2-CRY2 is different to that of the 158 kDa marker, indicating BIC2-CRY2 complex is a heterodimer with a 1:1 stoichiometry. Lower panel: an SDS-PAGE gel showing the proteins present in the peak fractions of BIC2-CRY2 from gel filtration chromatography visualized by Coomassie blue staining. c-d, SV-AUC and SLS analyses of the molecular weight of the full-length BIC2-CRY2 complex in solution under dark conditions. SLS analysis was performed in a SuperdexTM 200 increase 10/300 GL column. e, The domain of CRY2 interacting with BICs was characterized by pull-down assays. FL: full length; PHR: photolyase-homology region; CCE: Cryptochrome C-terminal Extension. Uncropped gel images of a, b and e are available as source data.
