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PubMed · 12454472

A medium-throughput crystallization approach.

Gerlind Sulzenbacher·Arnaud Gruez·Véronique Roig-Zamboni·Silvia Spinelli·Christel Valencia·Fabienne Pagot·Renaud Vincentelli·Christophe Bignon·Aurelia Salomoni·Sacha Grisel·Damien Maurin·Céline Huyghe·Kent Johansson·Alice Grassick·Alain Roussel·Yves Bourne·Sophie Perrier·Linda Miallau·Phillippe Cantau·Eric Blanc·Michel Genevois·Alain Grossi·André Zenatti·Valérie Campanacci·Christian Cambillau
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Abstract

The first results of a medium-scale structural genomics program clearly demonstrate the value of using a medium-throughput crystallization approach based on a two-step procedure: a large screening step employing robotics, followed by manual or automated optimization of the crystallization conditions. The structural genomics program was based on cloning in the Gateway vectors pDEST17, introducing a long 21-residue tail at the N-terminus. So far, this tail has not appeared to hamper crystallization. In ten months, 25 proteins were subjected to crystallization; 13 yielded crystals, of which ten led to usable data sets and five to structures. Furthermore, the results using a robot dispensing 50-200 nl drops indicate that smaller protein samples can be used for crystallization. These still partial results might indicate present and future directions for those who have to make crucial choices concerning their crystallization platform in structural genomics programs.

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CrystallizationEscherichia coli ProteinsLightRoboticsScattering, RadiationJournal ArticleResearch Support, Non-U.S. Gov't

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Gerlind Sulzenbacher, Arnaud Gruez, Véronique Roig-Zamboni, Silvia Spinelli, Christel Valencia, Fabienne Pagot, Renaud Vincentelli, Christophe Bignon, Aurelia Salomoni, Sacha Grisel, Damien Maurin, Céline Huyghe, Kent Johansson, Alice Grassick, Alain Roussel, Yves Bourne, Sophie Perrier, Linda Miallau, Phillippe Cantau, Eric Blanc, Michel Genevois, Alain Grossi, André Zenatti, Valérie Campanacci, Christian Cambillau. 2002-11-23. A medium-throughput crystallization approach.. https://doi.org/10.1107/s0907444902013938

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