Extended Data Fig. 9: SEC analyses of different CRY2 mutants.
From: Structural insights into the photoactivation of Arabidopsis CRY2
All the rans were performed in a SuperoseTM 6 increase 10/300 GL column. Left panel shows SEC analyses of CRY2WT (in darkness and under blue light), CRY2Y232F, CRY2Y232A, CRY2W353A and CRY2W353F. In darkness, wild type CRY2 eluted at 16.5 ml (deep teal line). Whereas, after blue light treatment, activated CRY2 eluted at 14.3 ml (dark purple line), indicating that CRY2 formed a tetramer. In darkness, CRY2Y232A (orange line) and a part of CRY2W353A (peak 1 of red line) eluted at around 14.8 ml, which is similar to that of CRY2WT under blue light, indicating CRY2Y232A and CRY2W353A are tetramers. CRY2Y232F (brown line) and CRY2W353F (green line) were eluted at 16.5 ml, indicating they were monomer. Right panel shows SEC analyses of CRY2WT (in darkness and under blue light), CRY2T244A, CRY2S245A, CRY2N356A and CRY2D387A. Coomassie blue-stained SDS-PAGE gels of peak fractions are on the right of corresponding SEC lines. Experiments were independently repeated three times with similar results. Uncropped gel images are available as source data.
