Compact, hydrophilic, lanthanide-binding tags for paramagnetic NMR spectroscopy

M D Lee, C T Loh, J Shin, S Chhabra, M L Dennis, G Otting, J D Swarbrick, B Graham

Research output: Contribution to journalArticleResearchpeer-review

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 Δχ-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.
Original languageEnglish
Pages (from-to)2614-2624
Number of pages11
JournalChemical Science
Volume6
Issue number4
DOIs
Publication statusPublished - 2015

Cite this

Lee, M. D., Loh, C. T., Shin, J., Chhabra, S., Dennis, M. L., Otting, G., ... Graham, B. (2015). Compact, hydrophilic, lanthanide-binding tags for paramagnetic NMR spectroscopy. Chemical Science, 6(4), 2614-2624. https://doi.org/10.1039/c4sc03892d
Lee, M D ; Loh, C T ; Shin, J ; Chhabra, S ; Dennis, M L ; Otting, G ; Swarbrick, J D ; Graham, B. / Compact, hydrophilic, lanthanide-binding tags for paramagnetic NMR spectroscopy. In: Chemical Science. 2015 ; Vol. 6, No. 4. pp. 2614-2624.
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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 Δχ-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.",
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Lee, MD, Loh, CT, Shin, J, Chhabra, S, Dennis, ML, Otting, G, Swarbrick, JD & Graham, B 2015, 'Compact, hydrophilic, lanthanide-binding tags for paramagnetic NMR spectroscopy' Chemical Science, vol. 6, no. 4, pp. 2614-2624. https://doi.org/10.1039/c4sc03892d

Compact, hydrophilic, lanthanide-binding tags for paramagnetic NMR spectroscopy. / Lee, M D; Loh, C T; Shin, J; Chhabra, S; Dennis, M L; Otting, G; Swarbrick, J D; Graham, B.

In: Chemical Science, Vol. 6, No. 4, 2015, p. 2614-2624.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Compact, hydrophilic, lanthanide-binding tags for paramagnetic NMR spectroscopy

AU - Lee, M D

AU - Loh, C T

AU - Shin, J

AU - Chhabra, S

AU - Dennis, M L

AU - Otting, G

AU - Swarbrick, J D

AU - Graham, B

PY - 2015

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AB - 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 Δχ-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.

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U2 - 10.1039/c4sc03892d

DO - 10.1039/c4sc03892d

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JO - Chemical Science

JF - Chemical Science

SN - 2041-6520

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Lee MD, Loh CT, Shin J, Chhabra S, Dennis ML, Otting G et al. Compact, hydrophilic, lanthanide-binding tags for paramagnetic NMR spectroscopy. Chemical Science. 2015;6(4):2614-2624. https://doi.org/10.1039/c4sc03892d