5XBO
Lanthanoid tagging via an unnatural amino acid for protein structure characterization
Summary for 5XBO
| Entry DOI | 10.2210/pdb5xbo/pdb |
| Descriptor | Polyubiquitin-B, UV excision repair protein RAD23 homolog A, TERBIUM(III) ION (3 entities in total) |
| Functional Keywords | unnatural amino acid, azide-alkyne cycloaddition, pseudo-contact shift, transient protein complex, protein binding |
| Biological source | Homo sapiens (Human) More |
| Cellular location | Ubiquitin: Cytoplasm : P0CG47 Nucleus: P54725 |
| Total number of polymer chains | 2 |
| Total formula weight | 14231.99 |
| Authors | |
| Primary citation | 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 Cited by 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: 28365903DOI: 10.1007/s10858-017-0106-9 PDB entries with the same primary citation |
| Experimental method | SOLUTION NMR |
Structure validation
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