4DNE

Crystal structure of a triple-mutant of streptavidin in complex with desthiobiotin

4DNE の概要

• エントリーDOI: 10.2210/pdb4dne/pdb
• 分子名称: Streptavidin, 6-(5-METHYL-2-OXO-IMIDAZOLIDIN-4-YL)-HEXANOIC ACID, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: biotin, biotin-binding protein
• 由来する生物種: Streptomyces avidinii
• 細胞内の位置: Secreted: P22629
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 38788.46

構造登録者

Panwar, P.,Deniaud, A.,Pebay-Peyroula, E. (登録日: 2012-02-08, 公開日: 2012-09-26, 最終更新日: 2023-09-13)

主引用文献

Panwar, P.,Deniaud, A.,Pebay-Peyroula, E.
Contamination from an affinity column: an encounter with a new villain in the world of membrane-protein crystallization.
Acta Crystallogr.,Sect.D, 68:1272-1277, 2012

PubMed Abstract: Attempts to crystallize AtNTT1, a chloroplast ATP/ADP transporter from Arabidopsis thaliana, revealed an unexpected contaminant, Strep-Tactin, a variant of streptavidin that was used during purification of the protein. Although it was present in very small amounts, crystals of Strep-Tactin were reproducibly grown from the AtNTT1 solution. AtNTT1 was overexpressed in Escherichia coli and purified from detergent-solubilized membrane fractions using Strep-Tactin affinity chromatography based on an engineered streptavidin. The contamination of protein solutions purified on Strep-Tactin columns has never been described previously and seems to be specific to membrane proteins solubilized in detergents. Trace amounts of Strep-Tactin were observed to be eluted from a Strep-Tactin column using several routinely used detergents, illustrating their possible role in the contamination. This finding raises an alarm and suggests caution in membrane-protein purification using Strep-Tactin affinity columns, where detergents are essential components. The small crystals of contaminant protein led to the structure at 1.9 Å resolution of Strep-Tactin in complex with desthiobiotin.

PubMed: 22993081
DOI: 10.1107/S090744491202639X

実験手法

X-RAY DIFFRACTION (1.88 Å)