2i5n

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Revision as of 10:02, 21 November 2007

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spinqualitylabelsall models 2i5n, resolution 1.96Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image

1.96 A X-ray structure of photosynthetic reaction center from Rhodopseudomonas viridis:Crystals grown by microfluidic technique

Overview

High-throughput screening and optimization experiments are critical to a, number of fields, including chemistry and structural and molecular, biology. The separation of these two steps may introduce false negatives, and a time delay between initial screening and subsequent optimization., Although a hybrid method combining both steps may address these problems, miniaturization is required to minimize sample consumption. This article, reports a "hybrid" droplet-based microfluidic approach that combines the, steps of screening and optimization into one simple experiment and uses, nanoliter-sized plugs to minimize sample consumption. Many distinct, reagents were sequentially introduced as approximately 140-nl plugs into a, microfluidic device and combined with a substrate and a diluting buffer., Tests were conducted in approximately 10-nl plugs containing different, concentrations of a reagent. Methods were developed to form plugs of, controlled concentrations, index concentrations, and incubate thousands of, plugs inexpensively and without evaporation. To validate the hybrid method, and demonstrate its applicability to challenging problems, crystallization, of model membrane proteins and handling of solutions of detergents and, viscous precipitants were demonstrated. By using 10 microl of protein, solution, approximately 1,300 crystallization trials were set up within 20, min by one researcher. This method was compatible with growth, manipulation, and extraction of high-quality crystals of membrane, proteins, demonstrated by obtaining high-resolution diffraction images and, solving a crystal structure. This robust method requires inexpensive, equipment and supplies, should be especially suitable for use in, individual laboratories, and could find applications in a number of areas, that require chemical, biochemical, and biological screening and, optimization.

About this Structure

2I5N is a Protein complex structure of sequences from Blastochloris viridis with FE2, SO4, HEC, BCB, BPB, MQ9, UQ1, NS5, LDA, HTO and UNL as ligands. Full crystallographic information is available from OCA.

Reference

Nanoliter microfluidic hybrid method for simultaneous screening and optimization validated with crystallization of membrane proteins., Li L, Mustafi D, Fu Q, Tereshko V, Chen DL, Tice JD, Ismagilov RF, Proc Natl Acad Sci U S A. 2006 Dec 19;103(51):19243-8. Epub 2006 Dec 11. PMID:17159147

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