SpringerProtocols: Abstract: How to Use Dynamic Light Scattering to Improve the Likelihood of Growing Macromolecular Crystals

How to Use Dynamic Light Scattering to Improve the Likelihood of Growing Macromolecular Crystals

By: Gloria E. O. Borgstahl²

Abstract

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Dynamic light scattering (DLS) has become one of the most useful diagnostic tools for crystallization. The main purpose of using DLS in crystal screening is to help the investigator understand the size distribution, stability, and aggregation state of macromolecules in solution. It can also be used to understand how experimental variables influence aggregation. With commercially available instruments, DLS is easy to perform, and most of the sample is recoverable. Most usefully, the homogeneity or monodispersity of a sample, as measured by DLS, can be predictive of crystallizability.

Affiliation(s): (2) Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE

Book Title: Macromolecular Crystallography Protocols: Volume 1, Preparation and Crystallization of Macromolecules

Series: Methods in Molecular Biology | Volume: 363 | Pub. Date: Feb-05-2007 | Page Range: 109-130 | DOI: 10.1007/978-1-59745-209-0_6

Subject: Protein Science

Key Words: Solubility - aggregation - hydrodynamic radius - monodispersity - polydispersity - dynamic light scattering - DLS

References

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1.	Zulauf, M. and D’Arcy, A. (1992) Light scattering of proteins as a criterion for crystallization. J. Cryst. Growth 122, 102–106. <Occurrence Type="DOI"><Handle>10.1016/0022-0248(92)90232-8</Handle></Occurrence>

2.	D’Arcy, A. (1994) Crystallizing proteins: a rational approach. Acta Cryst. D50, 469–471.

3.	Wilson, W. W. (2003) Light scattering as a diagnostic for protein crystal growth: a practical approach. J. Structural Biol. 142, 56–65. <Occurrence Type="DOI"><Handle>10.1016/S1047-8477(03)00038-8</Handle></Occurrence>

4.	Bergfors, T. M. (1999) Protein Crystallization Techniques, Strategies and, Tips. International University Line, La Jolla, CA.

5.	Ferré-D’Amaré, A. R. and Burley, S. K. (1997) Dynamic light scattering in evaluating crystallizability of macromolecules. Meth. Enzymol. 276, 157–166.

6.	Ranatunga, W., Jackson, D., Lloyd, J. A., Forget, A. L., Knight, K. L., and Borgstahl, G. E. O. (2001) Human Rad52 exhibits two modes of self-association. J. Biol. Chem. 276, 15,876–15,880.

7.	Mueser, T. C., Rogers, P. H., and Arnone, A. (2000) Interface sliding as illustrated by the multiple quaternary structures of liganded hemoglobin. Biochemistry 39, 15,353–15,364.

8.	Collins, B. K., Tomanicek, S. J., Lyamicheva, N., Kaiser, M. W., and Mueser, T. C. (2004) A preliminary solubility screen used to improve crystallization trials. Crystallization and preliminary X-ray structure determination of Aeropyrum pernix Flap Endonuclease-1. Acta Cryst. D60, 1674–1678.

9.	Jackson, D., Dhar, K., Wahl, J. K., Wold, M. S., and Borgstahl, G. E. (2002) Analysis of the human replication protein A:Rad52 complex: evidence for crosstalk between RPA32, RPA70, Rad52 and DNA. J. Mol. Biol. 321, 133–148. <Occurrence Type="DOI"><Handle>10.1016/S0022-2836(02)00541-7</Handle></Occurrence>

10.	Habel, J. E., Ohren, J. F., and Borgstahl, G. E. O. (2001) Dynamic light scattering analysis of full-length, human RPA14/32 dimer: purification, crystallization and self-association. Acta Cryst. D D57, 254–259.

11.	Dale, G. E., Oefner, C., and D’Arcy, A. (2003) The protein as a variable in protein crystallization. J. Struct. Biol. 142, 88–97. <Occurrence Type="DOI"><Handle>10.1016/S1047-8477(03)00041-8</Handle></Occurrence>

12.	Pusey, P. N., Koppel, D. E., Schaefer, D. W., Camerini-Otero, R. D., and Koenig, S. H. (1974) Intensity fluctuation spectroscopy of laser light scattered by solutions of spherical viruses: R17, Obeta, BSV, PM2 and T7. I. Light-scattering technique. Biochemistry 13, 952–959.

13.	Camerini-Otero, R. D., Pusey, P. N., Koppel, D. E., Schaefer, D. W., and Franklin, R. M. (1974) Intensity fluctuation spectroscopy of laser light scattered by solutions of spherical viruses: R17, Obeta, BSV, PM2 and T7. II. Diffusion coefficients, molecular weights, solvation, and particle dimensions. Biochemistry 13, 960–970.

14.	Pecora, R. (1985) Dynamic Light Scattering: Applications of Photon Correlation Spectroscopy. Plenum Press, New York, NY.

15.	Brown, R. G. W. (1990) Miniature laser light scattering instrumentation for particle size analysis. Applied Optics 29, 1.

16.	Phillies, G. D. J. (1990) Quasieleastic light scattering. Anal. Chem. 62, 1049A–1057A.

17.	Ranatunga, W., Jackson, D., II, R. A. F., and Borgstahl, G. E. O. (2001) Human Rad52 protein has extreme thermal stability. Biochemistry 40, 8557–8562.

18.	Long, M. M., Bishop, J. B., Nagabhushan, T. L., Reichert, P., Smith, G. D., and DeLucas, L. J. (1996) Protein crystal growth in microgravity: bovine insulin, human insulin and human alpha interferon. J. Crystal Growth 168, 233–243. <Occurrence Type="DOI"><Handle>10.1016/0022-0248(96)00325-9</Handle></Occurrence>

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