Compact, hydrophilic, lanthanide-binding tags for paramagnetic NMR spectroscopy.
Chem Sci. 2015 Apr 01;6(4):2614-2624
Authors: Lee MD, Loh CT, Shin J, Chhabra S, Dennis ML, Otting G, Swarbrick JD, Graham B
Abstract
The design, synthesis and evaluation of four novel lanthanide-binding tags for paramagnetic NMR spectroscopy are reported. Each tag is based on the ((2S,2'S,2''S,2'''S)-1,1',1'',1'''-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetrakis(propan-2-ol)) scaffold, featuring small chiral alcohol coordinating pendants to minimise the size and hydrophobic character of each tag. The tags feature different linkers of variable length for conjugation to protein via a single cysteine residue. Each tag's ability to induce pseudocontact shifts (PCS) was assessed on a ubiquitin A28C mutant. Two enantiomeric tags of particular note, C7 and C8, produced significantly larger ??-tensors compared to a previously developed tag, C1, attributed to the extremely short linker utilised, limiting the mobility of the bound lanthanide ion. The C7 and C8 tags' capacity to induce PCSs was further demonstrated on GB1 Q32C and 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK) S112C/C80A mutants. Whilst factors such as the choice of lanthanide ion, pH and site of conjugation influence the size of the PCSs obtained, the tags represent a significant advance in the field.
[NMR paper] Short two-armed lanthanide-binding tags for paramagnetic NMR spectroscopy based on chiral 1,4,7,10-tetrakis(2-hydroxypropyl)-1,4,7,10-tetraazacyclododecane scaffolds.
Short two-armed lanthanide-binding tags for paramagnetic NMR spectroscopy based on chiral 1,4,7,10-tetrakis(2-hydroxypropyl)-1,4,7,10-tetraazacyclododecane scaffolds.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.rsc.org-images-entities-char_z_RSClogo.gif Short two-armed lanthanide-binding tags for paramagnetic NMR spectroscopy based on chiral 1,4,7,10-tetrakis(2-hydroxypropyl)-1,4,7,10-tetraazacyclododecane scaffolds.
Chem Commun (Camb). 2017 Nov 22;:
Authors: Lee MD, Dennis ML, Graham B, Swarbrick JD
...
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[NMR paper] Enantiomeric two-armed lanthanide-binding tags for complementary effects in paramagnetic NMR spectroscopy.
Enantiomeric two-armed lanthanide-binding tags for complementary effects in paramagnetic NMR spectroscopy.
Related Articles Enantiomeric two-armed lanthanide-binding tags for complementary effects in paramagnetic NMR spectroscopy.
Chem Commun (Camb). 2016 Jun 2;
Authors: Lee MD, Dennis ML, Swarbrick JD, Graham B
Abstract
Two-armed lanthanide-binding tags induce significant, long-range paramagnetic effects in the NMR spectra of attached proteins. An enantiomeric pair of rigid, two-armed, cyclen-based tags are reported that...
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06-03-2016 04:52 PM
[NMR paper] Encoded loop-lanthanide-binding tags for long-range distance measurements in proteins by NMR and EPR spectroscopy.
Encoded loop-lanthanide-binding tags for long-range distance measurements in proteins by NMR and EPR spectroscopy.
Related Articles Encoded loop-lanthanide-binding tags for long-range distance measurements in proteins by NMR and EPR spectroscopy.
J Biomol NMR. 2015 Sep 4;
Authors: Barthelmes D, Gränz M, Barthelmes K, Allen KN, Imperiali B, Prisner T, Schwalbe H
Abstract
We recently engineered encodable lanthanide binding tags (LBTs) into proteins and demonstrated their applicability in Nuclear Magnetic Resonance (NMR)...
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09-06-2015 03:37 PM
Encoded loop-lanthanide-binding tags for long-range distance measurements in proteins by NMR and EPR spectroscopy
Encoded loop-lanthanide-binding tags for long-range distance measurements in proteins by NMR and EPR spectroscopy
Abstract
We recently engineered encodable lanthanide binding tags (LBTs) into proteins and demonstrated their applicability in Nuclear Magnetic Resonance (NMR) spectroscopy, X-ray crystallography and luminescence studies. Here, we engineered two-loop-LBTs into the model protein interleukin-1β (IL1β) and measured 1H, 15N-pseudocontact shifts (PCSs) by NMR spectroscopy. We determined the Î?Ï?-tensors associated with each Tm3+-loaded...
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09-05-2015 01:55 AM
[NMR paper] Protein structural studies by paramagnetic solid-state NMR spectroscopy aided by a compact cyclen-type Cu(II) binding tag.
Protein structural studies by paramagnetic solid-state NMR spectroscopy aided by a compact cyclen-type Cu(II) binding tag.
Protein structural studies by paramagnetic solid-state NMR spectroscopy aided by a compact cyclen-type Cu(II) binding tag.
J Biomol NMR. 2014 Nov 29;
Authors: Sengupta I, Gao M, Arachchige RJ, Nadaud PS, Cunningham TF, Saxena S, Schwieters CD, Jaroniec CP
Abstract
Paramagnetic relaxation enhancements (PREs) are a rich source of structural information in protein solid-state NMR spectroscopy. Here we...
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11-30-2014 09:41 PM
Protein structural studies by paramagnetic solid-state NMR spectroscopy aided by a compact cyclen-type Cu(II) binding tag
Protein structural studies by paramagnetic solid-state NMR spectroscopy aided by a compact cyclen-type Cu(II) binding tag
Abstract
Paramagnetic relaxation enhancements (PREs) are a rich source of structural information in protein solid-state NMR spectroscopy. Here we demonstrate that PRE measurements in natively diamagnetic proteins are facilitated by a thiol-reactive compact, cyclen-based, high-affinity Cu2+ binding tag, 1--1,4,7,10-tetraazacyclododecane (TETAC), that overcomes the key shortcomings associated with the use of larger, more flexible...
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11-29-2014 12:09 AM
An iminodiacetic acid based lanthanide binding tag for paramagnetic exchange NMR spectroscopy.
An iminodiacetic acid based lanthanide binding tag for paramagnetic exchange NMR spectroscopy.
An iminodiacetic acid based lanthanide binding tag for paramagnetic exchange NMR spectroscopy.
Angew Chem Int Ed Engl. 2011 May 2;50(19):4403-6
Authors: Swarbrick JD, Ung P, Chhabra S, Graham B
PMID: 21480444
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Engineering of a bis-chelator motif into a protein ?-helix for rigid lanthanide binding and paramagnetic NMR spectroscopy.
Engineering of a bis-chelator motif into a protein ?-helix for rigid lanthanide binding and paramagnetic NMR spectroscopy.
Engineering of a bis-chelator motif into a protein ?-helix for rigid lanthanide binding and paramagnetic NMR spectroscopy.
Chem Commun (Camb). 2011 May 27;
Authors: Swarbrick JD, Ung P, Su XC, Maleckis A, Chhabra S, Huber T, Otting G, Graham B
Attachment of two nitrilotriacetic acid-based ligands to a protein ?-helix in an i, i + 4 configuration produces an octadentate chelating motif that is able to bind paramagnetic...
Compact, hydrophilic, lanthanide-binding tags for paramagnetic NMR spectroscopy.
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