2I5N

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

2I5N の概要

• エントリーDOI: 10.2210/pdb2i5n/pdb
• 関連するPDBエントリー: 1DXR 1PRC
• 分子名称: Photosynthetic reaction center cytochrome c subunit, FE (II) ION, BACTERIOCHLOROPHYLL B, ... (16 entities in total)
• 機能のキーワード: photosynthetic reaction center, secondary quinone (qb), microfluidic technique, hybrid, microbatch, plug, photosynthesis
• 由来する生物種: Blastochloris viridis; 詳細
• 細胞内の位置: Cell membrane; Lipid-anchor: P07173; Cellular chromatophore membrane; Single-pass membrane protein: P06008; Cellular chromatophore membrane; Multi-pass membrane protein (By similarity): P06009 P06010
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 145662.27

構造登録者

Li, L.,Mustafi, D.,Fu, Q.,Tereshko, V.,Chen, D.L.,Tice, J.D.,Ismagilov, R.F. (登録日: 2006-08-25, 公開日: 2006-09-19, 最終更新日: 2024-11-20)

主引用文献

Li, L.,Mustafi, D.,Fu, Q.,Tereshko, V.,Chen, D.L.,Tice, J.D.,Ismagilov, R.F.
Nanoliter microfluidic hybrid method for simultaneous screening and optimization validated with crystallization of membrane proteins.
Proc.Natl.Acad.Sci.Usa, 103:19243-19248, 2006

PubMed Abstract: 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.

PubMed: 17159147
DOI: 10.1073/pnas.0607502103

実験手法

X-RAY DIFFRACTION (1.96 Å)