The plug-based nanovolume Microcapillary Protein Crystallization System (MPCS)
- Accelerated Technologies Center for Gene to 3D Structure (United States)
- deCODE biostructures Inc., 7869 NE Day Road West, Bainbridge Island, WA 98110 (United States)
- Department of Biochemistry, University of Washington, Seattle, WA 98195 (United States)
- Seattle Structural Genomics Center for Infectious Disease (United States)
- Emerald BioSystems Inc., 7869 NE Day Road West, Bainbridge Island, WA 98110 (United States)
The Microcapillary Protein Crystallization System (MPCS) is a new protein-crystallization technology used to generate nanolitre-sized crystallization experiments for crystal screening and optimization. Using the MPCS, diffraction-ready crystals were grown in the plastic MPCS CrystalCard and were used to solve the structure of methionine-R-sulfoxide reductase. The Microcapillary Protein Crystallization System (MPCS) embodies a new semi-automated plug-based crystallization technology which enables nanolitre-volume screening of crystallization conditions in a plasticware format that allows crystals to be easily removed for traditional cryoprotection and X-ray diffraction data collection. Protein crystals grown in these plastic devices can be directly subjected to in situ X-ray diffraction studies. The MPCS integrates the formulation of crystallization cocktails with the preparation of the crystallization experiments. Within microfluidic Teflon tubing or the microfluidic circuitry of a plastic CrystalCard, ∼10–20 nl volume droplets are generated, each representing a microbatch-style crystallization experiment with a different chemical composition. The entire protein sample is utilized in crystallization experiments. Sparse-matrix screening and chemical gradient screening can be combined in one comprehensive ‘hybrid’ crystallization trial. The technology lends itself well to optimization by high-granularity gradient screening using optimization reagents such as precipitation agents, ligands or cryoprotectants.
- OSTI ID:
- 22347989
- Journal Information:
- Acta Crystallographica. Section D: Biological Crystallography, Vol. 64, Issue Pt 11; Other Information: PMCID: PMC2585160; PUBLISHER-ID: bw5254; PMID: 19020349; OAI: oai:pubmedcentral.nih.gov:2585160; Copyright (c) Gerdts et al. 2008; This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.; Country of input: International Atomic Energy Agency (IAEA); ISSN 0907-4449
- Country of Publication:
- Denmark
- Language:
- English
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