5V33

R. sphaeroides photosythetic reaction center mutant - Residue L223, Ser to Trp - Room Temperature Structure Solved on X-ray Transparent Microfluidic Chip

5V33 の概要

• エントリーDOI: 10.2210/pdb5v33/pdb
• 分子名称: Reaction center protein H chain, Reaction center protein L chain, Reaction center protein M chain, ... (7 entities in total)
• 機能のキーワード: reaction center mutant, photosynthesis
• 由来する生物種: Rhodobacter sphaeroides; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 97683.96

構造登録者

Schieferstein, J.M.,Pawate, A.S.,Sun, C.,Wan, F.,Broecker, J.,Ernst, O.P.,Gennis, R.B.,Kenis, P.J.A. (登録日: 2017-03-06, 公開日: 2017-04-12, 最終更新日: 2023-10-04)

主引用文献

Schieferstein, J.M.,Pawate, A.S.,Sun, C.,Wan, F.,Sheraden, P.N.,Broecker, J.,Ernst, O.P.,Gennis, R.B.,Kenis, P.J.A.
X-ray transparent microfluidic chips for high-throughput screening and optimization of in meso membrane protein crystallization.
Biomicrofluidics, 11:024118-024118, 2017

PubMed Abstract: Elucidating and clarifying the function of membrane proteins ultimately requires atomic resolution structures as determined most commonly by X-ray crystallography. Many high impact membrane protein structures have resulted from advanced techniques such as crystallization that present technical difficulties for the set-up and scale-out of high-throughput crystallization experiments. In prior work, we designed a novel, low-throughput X-ray transparent microfluidic device that automated the mixing of protein and lipid by diffusion for crystallization trials. Here, we report X-ray transparent microfluidic devices for high-throughput crystallization screening and optimization that overcome the limitations of scale and demonstrate their application to the crystallization of several membrane proteins. Two complementary chips are presented: (1) a high-throughput screening chip to test 192 crystallization conditions in parallel using as little as 8 nl of membrane protein per well and (2) a crystallization optimization chip to rapidly optimize preliminary crystallization hits through fine-gradient re-screening. We screened three membrane proteins for new crystallization conditions, identifying several preliminary hits that we tested for X-ray diffraction quality. Further, we identified and optimized the crystallization condition for a photosynthetic reaction center mutant and solved its structure to a resolution of 3.5 Å.

PubMed: 28469762
DOI: 10.1063/1.4981818

実験手法

X-RAY DIFFRACTION (3.487 Å)