Optimization of crystallization of biological macromolecules using dialysis combined with temperature control

Junius, N.; Vahdatahar, E.; Oksanen, E.; Ferrer, J.-L.; Budayova-Spano, M.

doi:10.1107/S1600576720003209

Figure 8
The dialysis crystallization experiment with lactate dehydrogenase from T. thermophiles carried out at 293 K. (a) The beginning of the crystallization experiment (time 0). (b) Three days later, when the PEG 6000 concentration in the crystallization chamber is P1 = 2.5%(w/v). (c), (d) Two days after the PEG 6000 concentration was increased to P2 = 5%(w/v), crystals of the enzyme are observed and grow to large volume. (I) Situation observed in the comparative dialysis experiment carried out directly at P2 = 5% once the concentration difference between the compartments has been reached and (II) a few days later at equilibrium. (e) Schematic phase diagram (protein concentration versus precipitant concentration) incorporating selected images (to be tracked in ascending or alphabetical order) and illustrating the crystallization optimization workflow in growing large lactate dehydrogenase crystals. The enlargement of the metastable zone by reducing the rate generating the supersaturation is shown schematically. In accordance with a variant (not shown in Fig. 4

) of the standard workflow B1 (Section 2.3

), C1 represents the initial protein concentration used in the crystallization experiment (2.8 mg ml⁻¹) at P1 and C2 is the protein concentration corresponding to the relative equilibrium point reached at P2.

JOURNAL OF
APPLIED
CRYSTALLOGRAPHY

ISSN: 1600-5767

Volume 53| Part 3| June 2020| Pages 686-698

https://doi.org/10.1107/S1600576720003209

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