7BMS

HEWL in cesium chloride (1.5 M CsCl in crystallization condition and cryo protectant)

Summary for 7BMS

• Entry DOI: 10.2210/pdb7bms/pdb
• Descriptor: Lysozyme, CHLORIDE ION, CESIUM ION, ... (5 entities in total)
• Functional Keywords: cesium, phasing, hydrolase
• Biological source: Gallus gallus
• Total number of polymer chains: 1
• Total formula weight: 16661.52

Authors

Koelmel, W.,Kuper, J.,Kisker, C. (deposition date: 2021-01-20, release date: 2021-10-06, Last modification date: 2024-10-23)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.75 Å)