Journal
JOURNAL OF BIOMOLECULAR NMR
Volume 67, Issue 4, Pages 273-282Publisher
SPRINGER
DOI: 10.1007/s10858-017-0106-9
Keywords
Unnatural amino acid; Azide-alkyne cycloaddition; Pseudo-contact shift; Protein structure; Transient protein complex
Categories
Funding
- Chinese Ministry of Science and Technology [2013CB910200, 2016YFA0501200]
- National Natural Science Foundation of China [31225007, 31400644]
- K.C. Wong Education Foundation
- Howard Hughes Medical Institute
Ask authors/readers for more resources
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available