Figure 5 - full size

Figure 5.
A rationally designed, improved GCaMP2 variant. A, fluorescence excitation (solid lines) and emission (dashed lines) spectra of Ca^2+-saturated GCaMP2 T116V (red) and T116V/D381Y (blue), as well as their calcium-free forms (gray and black, respectively). Normalized absorbance spectra of each are shown in the inset. B, close-up stereo view of the Ca^2+-saturated monomeric GCaMP2 structure, showing the location of aspartate 381 (D381) of CaM at the CaM/cpEGFP domain interface. GCaMP2 is displayed as ribbons colored by domain. The side chain of Asp^381 and the cpEGFP chromophore are displayed as sticks. A model of a tyrosine side chain at position 381 is shown in semitransparent sticks to represent a possible conformation of the D381Y mutant and to illustrate the proximity of this side chain to the chromophore.