2OJR

Structure of ubiquitin solved by SAD using the Lanthanide-Binding Tag

2OJR の概要

• エントリーDOI: 10.2210/pdb2ojr/pdb
• 分子名称: Ubiquitin, TERBIUM(III) ION (3 entities in total)
• 機能のキーワード: lanthide-binding tag, terbium, tb, sad phasing, protein binding
• 由来する生物種: Homo sapiens (human)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 12761.65

構造登録者

Silvaggi, N.R.,Allen, K.N. (登録日: 2007-01-13, 公開日: 2007-06-12, 最終更新日: 2023-12-27)

主引用文献

Silvaggi, N.R.,Martin, L.J.,Schwalbe, H.,Imperiali, B.,Allen, K.N.
Double-Lanthanide-Binding Tags for Macromolecular Crystallographic Structure Determination.
J.Am.Chem.Soc., 129:7114-7120, 2007

PubMed Abstract: A double-lanthanide-binding tag (dLBT), a small peptide sequence engineered to bind two lanthanide ions (e.g., Tb3+) with high affinity, was used to solve the phase problem for the structure determination of ubiquitin by the single-wavelength anomalous diffraction (SAD) method. Since the dLBT is comprised exclusively of encoded amino acids, the necessity for the incorporation of unnatural amino acids or chemical modification of the protein as a prerequisite for X-ray structure determination is eliminated. A construct encoding the dLBT as an N-terminal fusion with ubiquitin provides for facile expression and purification using standard methods. Phasing of the single-wavelength X-ray data (at 2.6 A resolution) using only the anomalous signal from the two tightly bound Tb3+ ions in the dLBT led to clear electron-density maps. Nearly 75% of the ubiquitin structure was built using automated model-building software without user intervention. It is anticipated that this technique will be broadly applicable, complementing existing macromolecular phasing methodologies. The dLBT should be particularly useful in cases where protein derivatization with heavy atoms proves to be problematic or synchrotron facilities are unavailable.

PubMed: 17497863
DOI: 10.1021/ja070481n

実験手法

X-RAY DIFFRACTION (2.6 Å)