8FT6

The von Willebrand factor A domain of human capillary morphogenesis gene II, flexibly fused to the 1TEL crystallization chaperone, Ala-Ala linker variant, SUMO tag-free preparation.

8FT6 の概要

• エントリーDOI: 10.2210/pdb8ft6/pdb
• 関連するPDBエントリー: 8E1F 8FT8 8FZ3 8FZ4
• 分子名称: Transcription factor ETV6,Anthrax toxin receptor 2 chimera, CITRIC ACID, IODIDE ION, ... (5 entities in total)
• 機能のキーワード: telsam fusion, polymer forming crystallization chaperone, peptide binding protein
• 由来する生物種: Homo sapiens (human); 詳細
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 30514.09

構造登録者

Gajjar, P.L.,Litchfield, C.M.,Callahan, M.,Redd, N.,Doukov, T.,Moody, J.D. (登録日: 2023-01-11, 公開日: 2023-03-22, 最終更新日: 2023-10-18)

主引用文献

Gajjar, P.L.,Pedroza Romo, M.J.,Litchfield, C.M.,Callahan, M.,Redd, N.,Nawarathnage, S.,Soleimani, S.,Averett, J.,Wilson, E.,Lewis, A.,Stewart, C.,Tseng, Y.J.,Doukov, T.,Lebedev, A.,Moody, J.D.
Increasing the bulk of the 1TEL-target linker and retaining the 10×His tag in a 1TEL-CMG2-vWa construct improves crystal order and diffraction limits.
Acta Crystallogr D Struct Biol, 79:925-943, 2023

PubMed Abstract: TELSAM-fusion crystallization has the potential to become a revolutionary tool for the facile crystallization of proteins. TELSAM fusion can increase the crystallization rate and enable crystallization at low protein concentrations, in some cases with minimal crystal contacts [Nawarathnage et al. (2022), Open Biol. 12, 210271]. Here, requirements for the linker composition between 1TEL and a fused CMG2 vWa domain were investigated. Ala-Ala, Ala-Val, Thr-Val and Thr-Thr linkers were evaluated, comparing metrics for crystallization propensity and crystal order. The effect on crystallization of removing or retaining the purification tag was then tested. It was discovered that increasing the linker bulk and retaining the 10×His purification tag improved the diffraction resolution, likely by decreasing the number of possible vWa-domain orientations in the crystal. Additionally, it was discovered that some vWa-domain binding modes are correlated with scrambling of the 1TEL polymer orientation in crystals and an effective mitigation strategy for this pathology is presented.

PubMed: 37747038
DOI: 10.1107/S2059798323007246

実験手法

X-RAY DIFFRACTION (2.62 Å)