Semi-automatic protein crystallization system that allowsin situobservation of X-ray diffraction from crystals in the drop

2002 ◽  
Vol 58 (10) ◽  
pp. 1527-1530 ◽  
Author(s):  
Nobuhisa Watanabe ◽  
Hiroshi Murai ◽  
Isao Tanaka
Keyword(s):  
Protein Crystallization ◽  
X Ray Diffraction ◽  
X Ray ◽  
Crystallization System

scholarly journals Nanovolume optimization of protein crystal growth using the microcapillary protein crystallization system

2010 ◽  
Vol 43 (5) ◽  
pp. 1078-1083 ◽  
Author(s):  
Cory J. Gerdts ◽  
Glenn L. Stahl ◽  
Alberto Napuli ◽  
Bart Staker ◽  
Jan Abendroth ◽  
...  
Keyword(s):  
Protein Crystallization ◽  
Protein Structures ◽  
Protein Crystal ◽  
X Ray Diffraction ◽  
Vapor Diffusion ◽  
X Ray ◽  
Gradient Optimization ◽  
Crystallization System ◽  
Initial Crystal ◽  
Novel Protein

The Microcapillary Protein Crystallization System (MPCS) is a microfluidic, plug-based crystallization technology that generates X-ray diffraction-ready protein crystals in nanolitre volumes. In this study, 28 out of 29 (93%) proteins crystallized by traditional vapor diffusion experiments were successfully crystallized by chemical gradient optimization experiments using the MPCS technology. In total, 90 out of 120 (75%) protein/precipitant combinations leading to initial crystal hits from vapor diffusion experiments were successfully crystallized using MPCS technology. Many of the resulting crystals produced high-quality X-ray diffraction data, and six novel protein structures that were derived from crystals harvested from MPCS CrystalCards are reported.

Polysaccharides as Precipitants in Protein Crystallization for X-Ray Diffraction Studies

2011 ◽  
Vol 11 (4) ◽  
pp. 1152-1158 ◽  
Author(s):  
Laura Vepsäläinen ◽  
Katja Palmunen ◽  
Sinikka Uotila ◽  
Kalevi Visuri ◽  
Juha Rouvinen ◽  
...  
Keyword(s):  
Protein Crystallization ◽  
X Ray Diffraction ◽  
X Ray

scholarly journals In-plate protein crystallization,in situligand soaking and X-ray diffraction

2011 ◽  
Vol 67 (9) ◽  
pp. 747-755 ◽  
Author(s):  
Albane le Maire ◽  
Muriel Gelin ◽  
Sylvie Pochet ◽  
François Hoh ◽  
Michel Pirocchi ◽  
...  
Keyword(s):  
Protein Crystallization ◽  
X Ray Diffraction ◽  
X Ray

Combining `dry' co-crystallization andin situdiffraction to facilitate ligand screening by X-ray crystallography

2015 ◽  
Vol 71 (8) ◽  
pp. 1777-1787 ◽  
Author(s):  
Muriel Gelin ◽  
Vanessa Delfosse ◽  
Frédéric Allemand ◽  
François Hoh ◽  
Yoann Sallaz-Damaz ◽  
...  
Keyword(s):  
Protein Crystallization ◽  
X Ray Diffraction ◽  
X Ray ◽  
X Ray Crystallography ◽  
Ligand Screening ◽  
Alternative Approach ◽  
Manual Handling ◽  
Stock Solutions ◽  
Established Technique

X-ray crystallography is an established technique for ligand screening in fragment-based drug-design projects, but the required manual handling steps – soaking crystals with ligand and the subsequent harvesting – are tedious and limit the throughput of the process. Here, an alternative approach is reported: crystallization plates are pre-coated with potential binders prior to protein crystallization and X-ray diffraction is performed directly `in situ' (or in-plate). Its performance is demonstrated on distinct and relevant therapeutic targets currently being studied for ligand screening by X-ray crystallography using either a bending-magnet beamline or a rotating-anode generator. The possibility of using DMSO stock solutions of the ligands to be coated opens up a route to screening most chemical libraries.

Optimization of the critical nuclear size for protein crystallization: a note

2000 ◽  
Vol 56 (1) ◽  
pp. 106-108 ◽  
Author(s):  
Emmanuel Saridakis
Keyword(s):  
Local Minimum ◽  
Critical Radius ◽  
Protein Crystallization ◽  
Nuclear Size ◽  
X Ray Diffraction ◽  
X Ray

It was observed that for some proteins the best crystals for X-ray diffraction have been obtained at supersaturation ratios of ca 2.5–3 (in experiments without seeding). It was then noticed that under certain conditions specific to the protein such values are close to a local minimum of the critical radius for nucleation. A relation between the two observations is proposed.

On increasing protein-crystallization throughput for X-ray diffraction studies

2005 ◽  
Vol 61 (2) ◽  
pp. 123-129 ◽  
Author(s):  
Ashit K. Shah ◽  
Zhi-Jie Liu ◽  
Patrick D. Stewart ◽  
Florian D. Schubot ◽  
John P. Rose ◽  
...  
Keyword(s):  
Protein Crystallization ◽  
X Ray Diffraction ◽  
X Ray

scholarly journals Application of protein engineering to enhance crystallizability and improve crystal properties

2010 ◽  
Vol 66 (5) ◽  
pp. 604-615 ◽  
Author(s):  
Zygmunt S. Derewenda
Keyword(s):  
Protein Engineering ◽  
Model Building ◽  
Protein Crystallization ◽  
X Ray Diffraction ◽  
X Ray ◽  
Rate Limiting Step ◽  
Technological Advances ◽  
Radiation Sources ◽  
Crystal Properties ◽  
Automated Model Building

Until recently, protein crystallization has mostly been regarded as a stochastic event over which the investigator has little or no control. With the dramatic technological advances in synchrotron-radiation sources and detectors and the equally impressive progress in crystallographic software, including automated model building and validation, crystallization has increasingly become the rate-limiting step in X-ray diffraction studies of macromolecules. However, with the advent of recombinant methods it has also become possible to engineer target proteins and their complexes for higher propensity to form crystals with desirable X-ray diffraction qualities. As most proteins that are under investigation today are obtained by heterologous overexpression, these techniques hold the promise of becoming routine tools with the potential to transform classical crystallization screening into a more rational high-success-rate approach. This article presents an overview of protein-engineering methods designed to enhance crystallizability and discusses a number of examples of their successful application.

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