7BMX

p62PH in cesium chloride (0.25 M CsCl in protein buffer and 0.75 M CsCl in cryo protectant)

7BMX の概要

• エントリーDOI: 10.2210/pdb7bmx/pdb
• 分子名称: RNA polymerase II transcription factor B 73 kDa subunit-like protein, CESIUM ION, CHLORIDE ION, ... (4 entities in total)
• 機能のキーワード: cesium, phasing, pleckstrin homology domain, transcription
• 由来する生物種: Chaetomium thermophilum (strain DSM 1495 / CBS 144.50 / IMI 039719)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 16318.43

構造登録者

Koelmel, W.,Kuper, J.,Kisker, C. (登録日: 2021-01-20, 公開日: 2021-10-06, 最終更新日: 2024-06-19)

主引用文献

Koelmel, W.,Kuper, J.,Kisker, C.
Cesium based phasing of macromolecules: a general easy to use approach for solving the phase problem.
Sci Rep, 11:17038-17038, 2021

PubMed Abstract: Over the last decades the phase problem in macromolecular x-ray crystallography has become more controllable as methods and approaches have diversified and improved. However, solving the phase problem is still one of the biggest obstacles on the way of successfully determining a crystal structure. To overcome this caveat, we have utilized the anomalous scattering properties of the heavy alkali metal cesium. We investigated the introduction of cesium in form of cesium chloride during the three major steps of protein treatment in crystallography: purification, crystallization, and cryo-protection. We derived a step-wise procedure encompassing a "quick-soak"-only approach and a combined approach of CsCl supplement during purification and cryo-protection. This procedure was successfully applied on two different proteins: (i) Lysozyme and (ii) as a proof of principle, a construct consisting of the PH domain of the TFIIH subunit p62 from Chaetomium thermophilum for de novo structure determination. Usage of CsCl thus provides a versatile, general, easy to use, and low cost phasing strategy.

PubMed: 34426585
DOI: 10.1038/s41598-021-95186-1

実験手法

X-RAY DIFFRACTION (1.9 Å)