Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser

Wu, W.; Nogly, P.; Rheinberger, J.; Kick, L.M.; Gati, C.; Nelson, G.; Deupi, X.; Standfuss, J.; Schertler, G.; Panneels, V.

doi:10.1107/S2053230X15009966

Figure 1
Batch crystallization of rhodopsin. (a) Phase diagram representing the changes made to transform vapour-diffusion conditions to batch crystallization in order to obtain higher nucleation and micrometre-sized crystals of rhodopsin for SFX. The arrows in black and red show the routes to reach the nucleation and metastable zones during the crystallization process for the vapour-diffusion (VD) and batch methods, respectively. The insets show SHG images of rhodopsin crystals grown by vapour diffusion (left; long needles) and by batch (right; grain-shaped micrometre-sized crystals) (both insets represent a 200 µm square). The circled numbers refer to the phases `metastable zone' (1), `supersaturation zone' (2) and `precipitation zone' (3), which are successively reached when increasing the precipitant and/or protein concentration. (b) Three batch titrations of the precipitant into the rhodopsin solution under dim-light conditions. The upper and lower panels each show a scheme and a picture of an aliquot of an incomplete titration (tube 1; 20 µl rhodopsin + 32 µl precipitant) until low supersaturation [see label 1 in the phase diagram in (a)], an ideal batch titration (tube 2; 20 µl rhodopsin + 40 µl precipitant) until moderate supersaturation [see label 2 in the phase diagram in (a)] or an overtitration (tube 3; 20 µl rhodopsin + 50 µl precipitant) with an excess of precipitant inducing irreversible precipitation [see label 3 in the phase diagram in (

a)].

STRUCTURAL BIOLOGY
COMMUNICATIONS

ISSN: 2053-230X

Volume 71| Part 7| July 2015| Pages 856-860

https://doi.org/10.1107/S2053230X15009966

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