Optimization of protein crystallization: the OptiCryst project

Alfonso Garcia-Caballero, José A. Gavira, Estela Pineda-Molina, Naomi E. Chayen, Lata Govada, Khurshid A. Sheikh, Emmanuel Saridakis, Attia Boudjemline, Marcus J. Swann, Patrick Shaw Stewart, Richard A. Briggs, Stefan A. Kolek, Dominik Oberthuer, Karsten Dierks, Christian Betzel, Martha Santana, Jeanette R. Hobbs, Paul Thaw, Tony J. Savill, Jeroen R. MestersRolf Hilgenfeld, Nicklas Bonander, Roslyn Bill

Research output: Contribution to journalArticle

Abstract

Protein crystallization has gained a new strategic and commercial relevance in the postgenomic era due to its pivotal role in structural genomics. Producing high quality crystals has always been a bottleneck to efficient structure determination, and this problem is becoming increasingly acute. This is especially true for challenging, therapeutically important proteins that typically do not form suitable crystals. The OptiCryst consortium has focused on relieving this bottleneck by making a concerted effort to improve the crystallization techniques usually employed, designing new crystallization tools, and applying such developments to the optimization of target protein crystals. In particular, the focus has been on the novel application of dual polarization interferometry (DPI) to detect suitable nucleation; the application of in situ dynamic light scattering (DLS) to monitor and analyze the process of crystallization; the use of UV-fluorescence to differentiate protein crystals from salt; the design of novel nucleants and seeding technologies; and the development of kits for capillary counterdiffusion and crystal growth in gels. The consortium collectively handled 60 new target proteins that had not been crystallized previously. From these, we generated 39 crystals with improved diffraction properties. Fourteen of these 39 were only obtainable using OptiCryst methods. For the remaining 25, OptiCryst methods were used in combination with standard crystallization techniques. Eighteen structures have already been solved (30% success rate), with several more in the pipeline.
Original languageEnglish
Pages (from-to)2112–2121
Number of pages20
JournalCrystal Growth and Design
Volume11
Issue number6
DOIs
Publication statusPublished - 24 Mar 2011

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Crystallization
crystallization
proteins
Proteins
Crystals
optimization
crystals
relieving
kits
Dynamic light scattering
inoculation
Crystal growth
Interferometry
crystal growth
interferometry
Nucleation
light scattering
Gels
Pipelines
Diffraction

Cite this

Garcia-Caballero, A., Gavira, J. A., Pineda-Molina, E., Chayen, N. E., Govada, L., Sheikh, K. A., ... Bill, R. (2011). Optimization of protein crystallization: the OptiCryst project. Crystal Growth and Design, 11(6), 2112–2121. https://doi.org/10.1021/cg1013768
Garcia-Caballero, Alfonso ; Gavira, José A. ; Pineda-Molina, Estela ; Chayen, Naomi E. ; Govada, Lata ; Sheikh, Khurshid A. ; Saridakis, Emmanuel ; Boudjemline, Attia ; Swann, Marcus J. ; Shaw Stewart, Patrick ; Briggs, Richard A. ; Kolek, Stefan A. ; Oberthuer, Dominik ; Dierks, Karsten ; Betzel, Christian ; Santana, Martha ; Hobbs, Jeanette R. ; Thaw, Paul ; Savill, Tony J. ; Mesters, Jeroen R. ; Hilgenfeld, Rolf ; Bonander, Nicklas ; Bill, Roslyn. / Optimization of protein crystallization : the OptiCryst project. In: Crystal Growth and Design. 2011 ; Vol. 11, No. 6. pp. 2112–2121.
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Garcia-Caballero, A, Gavira, JA, Pineda-Molina, E, Chayen, NE, Govada, L, Sheikh, KA, Saridakis, E, Boudjemline, A, Swann, MJ, Shaw Stewart, P, Briggs, RA, Kolek, SA, Oberthuer, D, Dierks, K, Betzel, C, Santana, M, Hobbs, JR, Thaw, P, Savill, TJ, Mesters, JR, Hilgenfeld, R, Bonander, N & Bill, R 2011, 'Optimization of protein crystallization: the OptiCryst project', Crystal Growth and Design, vol. 11, no. 6, pp. 2112–2121. https://doi.org/10.1021/cg1013768

Optimization of protein crystallization : the OptiCryst project. / Garcia-Caballero, Alfonso; Gavira, José A.; Pineda-Molina, Estela; Chayen, Naomi E.; Govada, Lata; Sheikh, Khurshid A.; Saridakis, Emmanuel; Boudjemline, Attia; Swann, Marcus J.; Shaw Stewart, Patrick; Briggs, Richard A.; Kolek, Stefan A.; Oberthuer, Dominik; Dierks, Karsten; Betzel, Christian; Santana, Martha; Hobbs, Jeanette R.; Thaw, Paul; Savill, Tony J.; Mesters, Jeroen R.; Hilgenfeld, Rolf; Bonander, Nicklas; Bill, Roslyn.

In: Crystal Growth and Design, Vol. 11, No. 6, 24.03.2011, p. 2112–2121.

Research output: Contribution to journalArticle

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T1 - Optimization of protein crystallization

T2 - the OptiCryst project

AU - Garcia-Caballero, Alfonso

AU - Gavira, José A.

AU - Pineda-Molina, Estela

AU - Chayen, Naomi E.

AU - Govada, Lata

AU - Sheikh, Khurshid A.

AU - Saridakis, Emmanuel

AU - Boudjemline, Attia

AU - Swann, Marcus J.

AU - Shaw Stewart, Patrick

AU - Briggs, Richard A.

AU - Kolek, Stefan A.

AU - Oberthuer, Dominik

AU - Dierks, Karsten

AU - Betzel, Christian

AU - Santana, Martha

AU - Hobbs, Jeanette R.

AU - Thaw, Paul

AU - Savill, Tony J.

AU - Mesters, Jeroen R.

AU - Hilgenfeld, Rolf

AU - Bonander, Nicklas

AU - Bill, Roslyn

PY - 2011/3/24

Y1 - 2011/3/24

N2 - Protein crystallization has gained a new strategic and commercial relevance in the postgenomic era due to its pivotal role in structural genomics. Producing high quality crystals has always been a bottleneck to efficient structure determination, and this problem is becoming increasingly acute. This is especially true for challenging, therapeutically important proteins that typically do not form suitable crystals. The OptiCryst consortium has focused on relieving this bottleneck by making a concerted effort to improve the crystallization techniques usually employed, designing new crystallization tools, and applying such developments to the optimization of target protein crystals. In particular, the focus has been on the novel application of dual polarization interferometry (DPI) to detect suitable nucleation; the application of in situ dynamic light scattering (DLS) to monitor and analyze the process of crystallization; the use of UV-fluorescence to differentiate protein crystals from salt; the design of novel nucleants and seeding technologies; and the development of kits for capillary counterdiffusion and crystal growth in gels. The consortium collectively handled 60 new target proteins that had not been crystallized previously. From these, we generated 39 crystals with improved diffraction properties. Fourteen of these 39 were only obtainable using OptiCryst methods. For the remaining 25, OptiCryst methods were used in combination with standard crystallization techniques. Eighteen structures have already been solved (30% success rate), with several more in the pipeline.

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Garcia-Caballero A, Gavira JA, Pineda-Molina E, Chayen NE, Govada L, Sheikh KA et al. Optimization of protein crystallization: the OptiCryst project. Crystal Growth and Design. 2011 Mar 24;11(6):2112–2121. https://doi.org/10.1021/cg1013768