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


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.

SUBMITTER: Lee MD 

PROVIDER: S-EPMC5812434 | biostudies-literature | 2015 Apr

REPOSITORIES: biostudies-literature

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

Lee M D MD   Loh C-T CT   Shin J J   Chhabra S S   Dennis M L ML   Otting G G   Swarbrick J D JD   Graham B B  

Chemical science 20150225 4


The design, synthesis and evaluation of four novel lanthanide-binding tags for paramagnetic NMR spectroscopy are reported. Each tag is based on the ((2<i>S</i>,2'<i>S</i>,2''<i>S</i>,2'''<i>S</i>)-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 <i>via</i> a  ...[more]

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