Efficient Cryoprotection of Macromolecular Crystals Using Vapor Diffusion of Volatile Alcohols

Overview

Journal J Struct Biol

Specialties Cell Biology
Molecular Biology

Date 2014 Oct 7

PMID 25286441

Citations 8

Authors

Christopher Farley

Douglas H Juers

Affiliations

Soon will be listed here.

Abstract

Macromolecular X-ray crystallography, usually done at cryogenic temperature to limit radiation damage, often requires liquid cryoprotective soaking that can be labor intensive and damaging to crystals. Here we describe a method for cryoprotection that uses vapor diffusion of volatile cryoprotective agents into loop-mounted crystals. The crystal is mounted into a vial containing a small volume of an alcohol-based cryosolution. After a short incubation with the looped crystal sitting in the cryosolution vapor, the crystal is transferred directly from the vial into the cooling medium. Effective for several different protein crystals, the approach obviates the need for liquid soaking and opens up a heretofore underutilized class of cryoprotective agents for macromolecular crystallography.

Citing Articles

A drug-discovery-oriented non-invasive protocol for protein crystal cryoprotection by dehydration, with application for crystallization screening.

Bellini D J Appl Crystallogr. 2022; 55(Pt 2):370-379.

PMID: 35497658 PMC: 8985602. DOI: 10.1107/S1600576722002382.

Alpha-Lipoic Acid Can Overcome The Reduced Developmental Competency Induced by Alcohol Toxicity during Ovine Oocyte Maturation.

Moghimi Khorasgani A, Moradi R, Jafarpour F, Ghazvinizadehgan F, Ostadhosseini S, Heydarnezhad A Cell J. 2021; 23(2):164-173.

PMID: 34096217 PMC: 8196229. DOI: 10.22074/cellj.2021.7071.

Structural Insights into the Recognition of Mono- and Diacetylated Histones by the ATAD2B Bromodomain.

Lloyd J, McLaughlin K, Lubula M, Gay J, Dest A, Gao C J Med Chem. 2020; 63(21):12799-12813.

PMID: 33084328 PMC: 7884259. DOI: 10.1021/acs.jmedchem.0c01178.

A comparison of gas stream cooling and plunge cooling of macromolecular crystals.

Harrison K, Wu Z, Juers D J Appl Crystallogr. 2019; 52(Pt 5):1222-1232.

PMID: 31636524 PMC: 6782077. DOI: 10.1107/S1600576719010318.

The impact of cryosolution thermal contraction on proteins and protein crystals: volumes, conformation and order.

Juers D, Farley C, Saxby C, Cotter R, Cahn J, Holton-Burke R Acta Crystallogr D Struct Biol. 2018; 74(Pt 9):922-938.

PMID: 30198901 PMC: 6130464. DOI: 10.1107/S2059798318008793.

References

Kim C, Kapfer R, Gruner S . High-pressure cooling of protein crystals without cryoprotectants. Acta Crystallogr D Biol Crystallogr. 2005; 61(Pt 7):881-90. DOI: 10.1107/S090744490500836X. View

Auton M, Rosgen J, Sinev M, Holthauzen L, Bolen D . Osmolyte effects on protein stability and solubility: a balancing act between backbone and side-chains. Biophys Chem. 2011; 159(1):90-9. PMC: 3166983. DOI: 10.1016/j.bpc.2011.05.012. View

Juers D, Matthews B . Reversible lattice repacking illustrates the temperature dependence of macromolecular interactions. J Mol Biol. 2001; 311(4):851-62. DOI: 10.1006/jmbi.2001.4891. View

Berman H, Westbrook J, Feng Z, Gilliland G, Bhat T, Weissig H . The Protein Data Bank. Nucleic Acids Res. 1999; 28(1):235-42. PMC: 102472. DOI: 10.1093/nar/28.1.235. View

Hausrath A, Matthews B . Thermolysin in the absence of substrate has an open conformation. Acta Crystallogr D Biol Crystallogr. 2002; 58(Pt 6 Pt 2):1002-7. DOI: 10.1107/s090744490200584x. View

Pemberton T, Still B, Christensen E, Singh H, Srivastava D, Tanner J . Proline: Mother Nature's cryoprotectant applied to protein crystallography. Acta Crystallogr D Biol Crystallogr. 2012; 68(Pt 8):1010-8. PMC: 3413213. DOI: 10.1107/S0907444912019580. View

Leiros H, McSweeney S, Smalas A . Atomic resolution structures of trypsin provide insight into structural radiation damage. Acta Crystallogr D Biol Crystallogr. 2001; 57(Pt 4):488-97. DOI: 10.1107/s0907444901000646. View

Kantardjieff K, Rupp B . Matthews coefficient probabilities: Improved estimates for unit cell contents of proteins, DNA, and protein-nucleic acid complex crystals. Protein Sci. 2003; 12(9):1865-71. PMC: 2323984. DOI: 10.1110/ps.0350503. View

Low B, Chen C, Berger J, Singman L, Pletcher J . Studies of insulin crystals at low temperatures: effects on mosaic character and radiation sensitivity. Proc Natl Acad Sci U S A. 1966; 56(6):1746-50. PMC: 220166. DOI: 10.1073/pnas.56.6.1746. View

10.

Warkentin M, Sethna J, Thorne R . Critical droplet theory explains the glass formability of aqueous solutions. Phys Rev Lett. 2013; 110(1):015703. DOI: 10.1103/PhysRevLett.110.015703. View

11.

Juers D, Matthews B . Cryo-cooling in macromolecular crystallography: advantages, disadvantages and optimization. Q Rev Biophys. 2005; 37(2):105-19. DOI: 10.1017/s0033583504004007. View

12.

Burkhardt A, Warmer M, Panneerselvam S, Wagner A, Zouni A, Glockner C . Fast high-pressure freezing of protein crystals in their mother liquor. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012; 68(Pt 4):495-500. PMC: 3325829. DOI: 10.1107/S1744309112009670. View

13.

Matthews B . Solvent content of protein crystals. J Mol Biol. 1968; 33(2):491-7. DOI: 10.1016/0022-2836(68)90205-2. View

14.

Alcorn T, Juers D . Progress in rational methods of cryoprotection in macromolecular crystallography. Acta Crystallogr D Biol Crystallogr. 2010; 66(Pt 4):366-73. PMC: 2852300. DOI: 10.1107/S090744490903995X. View

15.

Kmetko J, Husseini N, Naides M, Kalinin Y, Thorne R . Quantifying X-ray radiation damage in protein crystals at cryogenic temperatures. Acta Crystallogr D Biol Crystallogr. 2006; 62(Pt 9):1030-8. DOI: 10.1107/S0907444906023869. View

16.

Kriminski S, Caylor C, Nonato M, Finkelstein K, Thorne R . Flash-cooling and annealing of protein crystals. Acta Crystallogr D Biol Crystallogr. 2002; 58(Pt 3):459-71. DOI: 10.1107/s0907444902000112. View

17.

Douzou P, Hoa G, Petsko G . Protein crystallography at sub-zero temperatures: lysozyme-substrate complexes in cooled mixed solvents. J Mol Biol. 1975; 96(3):367-80. DOI: 10.1016/0022-2836(75)90166-7. View

18.

Russi S, Juers D, Sanchez-Weatherby J, Pellegrini E, Mossou E, Forsyth V . Inducing phase changes in crystals of macromolecules: status and perspectives for controlled crystal dehydration. J Struct Biol. 2011; 175(2):236-43. DOI: 10.1016/j.jsb.2011.03.002. View

19.

Rubinson K, Ladner J, Tordova M, Gilliland G . Cryosalts: suppression of ice formation in macromolecular crystallography. Acta Crystallogr D Biol Crystallogr. 2000; 56(Pt 8):996-1001. DOI: 10.1107/s0907444900007587. View

20.

Bujacz G, Wrzesniewska B, Bujacz A . Cryoprotection properties of salts of organic acids: a case study for a tetragonal crystal of HEW lysozyme. Acta Crystallogr D Biol Crystallogr. 2010; 66(Pt 7):789-96. DOI: 10.1107/S0907444910015416. View