Thermal contraction of aqueous glycerol and ethylene glycol solutions for optimized protein-crystal cryoprotection.
The thermal contraction of aqueous cryoprotectant solutions on cooling to cryogenic temperatures is of practical importance in protein cryocrystallography and in biological cryopreservation. In the former case, differential contraction on cooling of protein molecules and their lattice relative to that of the internal and surrounding solvent may lead to crystal damage and the degradation of crystal diffraction properties. Here, the amorphous phase densities of aqueous solutions of glycerol and ethylene glycol at T = 77 K have been determined. Densities with accuracies of <0.5% to concentrations as low as 30%(w/v) were determined by rapidly cooling drops with volumes as small as 70 pl, assessing their optical clarity and measuring their buoyancy in liquid nitrogen-argon solutions. The use of these densities in contraction matching of internal solvent to the available solvent spaces is complicated by several factors, most notably the exclusion of cryoprotectants from protein hydration shells and the expected deviation of the contraction behavior of hydration water from bulk water. The present methods and results will assist in developing rational approaches to cryoprotection and an understanding of solvent behavior in protein crystals.
Cornell University, Ithaca, NY 14853, USA.
Chen
C
Shen
Cornell University, Ithaca, NY 14853, USA.
Ethan F
EF
Julius
Cornell University, Ithaca, NY 14853, USA.
Timothy J
TJ
Tyree
Physics Department, Cornell University, Ithaca, NY 14853, USA.
David W
DW
Moreau
Physics Department, Cornell University, Ithaca, NY 14853, USA.
Hakan
H
Atakisi
Physics Department, Cornell University, Ithaca, NY 14853, USA.
Robert E
RE
Thorne
D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods*; D003451:Cryoprotective Agents / Q000737:chemistry*; D004867:Equipment Design; D019855:Ethylene Glycol / Q000737:chemistry*; D005990:Glycerol / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D018401:Sample Size; D013696:Temperature; D058989:Vitrification; D014867:Water / Q000737:chemistry
D003451:Cryoprotective Agents; D011506:Proteins; D014867:Water; D019855:Ethylene Glycol; D005990:Glycerol
aqueous solution; buoyancy; cryocrystallography; cryoflotation; cryopreservation; density; expansion matching; protein crystallography; thermal expansion; vitreous; vitrification