6FJS

Proteinase~K SIRAS phased structure of room-temperature, serially collected synchrotron data

6FJS の概要

• エントリーDOI: 10.2210/pdb6fjs/pdb
• 分子名称: Proteinase K, CALCIUM ION (3 entities in total)
• 機能のキーワード: hydrolase, protease, serine protease
• 由来する生物種: Parengyodontium album (Tritirachium album)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 29014.76

構造登録者

Botha, S.,Baitan, D.,Jungnickel, K.E.J.,Oberthuer, D.,Schmidt, C.,Stern, S.,Wiedorn, M.O.,Perbandt, M.,Chapman, H.N.,Betzel, C. (登録日: 2018-01-23, 公開日: 2018-10-10)

主引用文献

Botha, S.,Baitan, D.,Jungnickel, K.E.J.,Oberthur, D.,Schmidt, C.,Stern, S.,Wiedorn, M.O.,Perbandt, M.,Chapman, H.N.,Betzel, C.
De novoprotein structure determination by heavy-atom soaking in lipidic cubic phase and SIRAS phasing using serial synchrotron crystallography.
IUCrJ, 5:524-530, 2018

PubMed Abstract: During the past few years, serial crystallography methods have undergone continuous development and serial data collection has become well established at high-intensity synchrotron-radiation beamlines and XFEL radiation sources. However, the application of experimental phasing to serial crystallography data has remained a challenging task owing to the inherent inaccuracy of the diffraction data. Here, a particularly gentle method for incorporating heavy atoms into micrometre-sized crystals utilizing lipidic cubic phase (LCP) as a carrier medium is reported. Soaking in LCP prior to data collection offers a new, efficient and gentle approach for preparing heavy-atom-derivative crystals directly before diffraction data collection using serial crystallography methods. This approach supports effective phasing by utilizing a reasonably low number of diffraction patterns. Using synchrotron radiation and exploiting the anomalous scattering signal of mercury for single isomorphous replacement with anomalous scattering (SIRAS) phasing resulted in high-quality electron-density maps that were sufficient for building a complete structural model of proteinase K at 1.9 Å resolution using automatic model-building tools.

PubMed: 30224955
DOI: 10.1107/S2052252518009223

実験手法

X-RAY DIFFRACTION (1.9 Å)