Site-specific tagging proteins with a rigid, small and stable transition metal chelator, 8-hydroxyquinoline, for paramagnetic NMR analysis
-
Altmetric Citations
Yang, Yin; Huang, Feng; Huber, Thomas; Su, Xun-Cheng
Description
Design of a paramagnetic metal binding motif in a protein is a valuable way for understanding the function, dynamics and interactions of a protein by paramagnetic NMR spectroscopy. Several strategies have been proposed to site-specifically tag proteins with paramagnetic lanthanide ions. Here we report a simple approach of engineering a transition metal binding motif via site-specific labelling of a protein with 2-vinyl-8-hydroxyquinoline (2V-8HQ). The protein-2V-8HQ adduct forms a stable...[Show more]
dc.contributor.author | Yang, Yin | |
---|---|---|
dc.contributor.author | Huang, Feng | |
dc.contributor.author | Huber, Thomas | |
dc.contributor.author | Su, Xun-Cheng | |
dc.date.accessioned | 2018-11-13T04:16:05Z | |
dc.date.available | 2018-11-13T04:16:05Z | |
dc.identifier.issn | 0925-2738 | |
dc.identifier.uri | http://hdl.handle.net/1885/149437 | |
dc.description.abstract | Design of a paramagnetic metal binding motif in a protein is a valuable way for understanding the function, dynamics and interactions of a protein by paramagnetic NMR spectroscopy. Several strategies have been proposed to site-specifically tag proteins with paramagnetic lanthanide ions. Here we report a simple approach of engineering a transition metal binding motif via site-specific labelling of a protein with 2-vinyl-8-hydroxyquinoline (2V-8HQ). The protein-2V-8HQ adduct forms a stable complex with transition metal ions, Mn(II), Co(II), Ni(II), Cu(II) and Zn(II). The paramagnetic effects generated by these transition metal ions were evaluated by NMR spectroscopy. We show that 2V-8HQ is a rigid and stable transition metal binding tag. The coordination of the metal ion can be assisted by protein sidechains. More importantly, tunable paramagnetic tensors are simply obtained in an α-helix that possesses solvent exposed residues in positions i and i + 3, where i is the residue to be mutated to cysteine, i + 3 is Gln or Glu or i - 4 is His. The coordination of a sidechain carboxylate/amide or imidazole to cobalt(II) results in different structural geometries, leading to different paramagnetic tensors as shown by experimental data. | |
dc.description.sponsorship | Financial support by the 973 program (2013CB910200), the National Science Foundation of China (21473095 and 21273121), and the Australian Research Council (DP120100561 and DP150100383) is greatly acknowledged. | |
dc.format.mimetype | application/pdf | |
dc.source | Journal of biomolecular NMR | |
dc.subject | 8-hydroxyquinoline | |
dc.subject | paramagnetic nmr spectroscopy | |
dc.subject | paramagnetic relaxation enhancement | |
dc.subject | protein labeling | |
dc.subject | pseudocontact shift | |
dc.subject | animals | |
dc.subject | chelating agents | |
dc.subject | humans | |
dc.subject | oxyquinoline | |
dc.subject | ubiquitin | |
dc.subject | nuclear magnetic resonance, biomolecular | |
dc.title | Site-specific tagging proteins with a rigid, small and stable transition metal chelator, 8-hydroxyquinoline, for paramagnetic NMR analysis | |
dc.type | Journal article | |
local.identifier.citationvolume | 64 | |
dc.date.issued | 2016-02 | |
local.type.status | Accepted Version | |
dc.relation | http://purl.org/au-research/grants/arc/DP120100561 | |
dc.relation | http://purl.org/au-research/grants/arc/DP150100383 | |
local.identifier.essn | 1573-5001 | |
local.bibliographicCitation.issue | 2 | |
local.bibliographicCitation.startpage | 103-13 | |
local.bibliographicCitation.lastpage | 113 | |
local.identifier.doi | 10.1007/s10858-016-0011-7 | |
dcterms.accessRights | Open Access | |
dc.provenance | Author can archive post-print (ie final draft post-refereeing) | |
Collections | ANU Research Publications |
Download
File | Description | Size | Format | Image |
---|---|---|---|---|
Yang_et_al.pdf | Author/s Accepted Manuscript (AAM) / Post-print | 1.22 MB | Adobe PDF |
Items in Open Research are protected by copyright, with all rights reserved, unless otherwise indicated.
Updated: 17 November 2022/ Responsible Officer: University Librarian/ Page Contact: Library Systems & Web Coordinator