Fig. 1
From: Understanding CRY2 interactions for optical control of intracellular signaling
Electrostatic charges at N- and C-termini are important for CRY2–CIB1 and CRY2–CRY2 interactions, respectively. a Surface charge distribution of CRY2wt predicted by homology modeling. Charged residues are labeled in the insets of both the N-terminus and C-terminus. b Electrostatic charges at the N-terminus of CRY2 are removed by having charged amino acids replaced with neutral ones (CRY2(neutral2–6)) or deleted (CRY2(Δ2–6)). c The light-induced CRY2–CIB1 binding is much weaker for CRY2(neutral2-6) or CRY2(Δ2–6), as compared to CRY2wt. COS7 cells were co-transfected with CIB1-GFP-Sec61 and each mCh-tagged CRY2, respectively. Blue light was delivered at 2-s intervals for 100 s. d The amino acid sequences for the C-terminus of CRY2wt, CRY2(E490G) and each truncated CRY2. Truncated CRY2 variants, including CRY2(Δ491–498), CRY2(Δ490–498), CRY2(Δ489–498), CRY2(Δ488–498) and CRY2(Δ487–498), were constructed by sequentially removing residues from CRY2(487–490). e Cytosolic CRY2(E490G) and CRY2(Δ490–498) formed clusters upon light stimulation, while the other C-terminally truncated derivatives did not. COS7 cells were transfected with each mCh-tagged CRY2 and stimulated with blue light at 5-s intervals for 500 s. Scale bars, 5 µm (c), 10 µm (e)
