5XBO

Lanthanoid tagging via an unnatural amino acid for protein structure characterization

5XBO の概要

• エントリーDOI: 10.2210/pdb5xbo/pdb
• 分子名称: Polyubiquitin-B, UV excision repair protein RAD23 homolog A, TERBIUM(III) ION (3 entities in total)
• 機能のキーワード: unnatural amino acid, azide-alkyne cycloaddition, pseudo-contact shift, transient protein complex, protein binding
• 由来する生物種: Homo sapiens (Human); 詳細
• 細胞内の位置: Ubiquitin: Cytoplasm : P0CG47; Nucleus: P54725
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 14231.99

構造登録者

Jiang, W.,Gu, X.,Dong, X.,Tang, C. (登録日: 2017-03-21, 公開日: 2017-05-31, 最終更新日: 2024-05-15)

主引用文献

Jiang, W.X.,Gu, X.H.,Dong, X.,Tang, C.
Lanthanoid tagging via an unnatural amino acid for protein structure characterization
J. Biomol. NMR, 67:273-282, 2017

PubMed Abstract: Lanthanoid pseudo-contact shift (PCS) provides long-range structural information between a paramagnetic tag and protein nuclei. However, for proteins with native cysteines, site-specific attachment may only utilize functional groups orthogonal to sulfhydryl chemistry. Here we report two lanthanoid probes, DTTA-C3-yne and DTTA-C4-yne, which can be conjugated to an unnatural amino acid pAzF in the target protein via azide-alkyne cycloaddition. Demonstrated with ubiquitin and cysteine-containing enzyme EIIB, we show that large PCSs of distinct profiles can be generated for each tag/lanthanoid combination. The DTTA-based lanthanoid tags are associated with large magnetic susceptibility tensors owing to the rigidity of the tags. In particular, introduction of the DTTA-C3 tag affords intermolecular PCSs and enables structural characterization of a transient protein complex between ubiquitin and a UBA domain. Together, we have expanded the repertoire of paramagnetic tags and the applicability of paramagnetic NMR.

PubMed: 28365903
DOI: 10.1007/s10858-017-0106-9

実験手法

SOLUTION NMR