Cys-Ph-TAHA: a lanthanide binding tag for RDC and PCS enhanced protein NMR

被引：35

作者：

Peters, Fabian ^{[1
]}

Maestre-Martinez, Mitcheell ^{[1
]}

Leonov, Andrei ^{[1
]}

Kovacic, Lidija ^{[2
,3
]}

Becker, Stefan ^{[1
]}

Boelens, Rolf ^{[2
]}

Griesinger, Christian ^{[1
]}

机构：

[1] Max Planck Inst Biophys Chem, Dept NMR Based Struct Biol, D-37077 Gottingen, Germany

[2] Univ Utrecht, Dept NMR Spect, Bijvoet Ctr Biomol Res, NL-3584 CH Utrecht, Netherlands

[3] Jozef Stefan Inst, Dept Mol & Biomed Sci, Ljubljana 1000, Slovenia

来源：

JOURNAL OF BIOMOLECULAR NMR | 2011年 / 51卷 / 03期

关键词：

Lanthanides; Pseudocontact shifts; Residual dipolar couplings; Paramagnetic NMR; RESIDUAL DIPOLAR COUPLINGS; MAGNETIC-SUSCEPTIBILITY; PARAMAGNETIC NMR; WEAK ALIGNMENT; UBIQUITIN; DYNAMICS; DESIGN; ACID; RECOGNITION; EXPRESSION;

D O I：

10.1007/s10858-011-9560-y

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Here we present Cys-Ph-TAHA, a new nonadentate lanthanide tag for the paramagnetic labelling of proteins. The tag can be easily synthesized and is stereochemically homogenous over a wide range of temperatures, yielding NMR spectra with a single set of peaks. Bound to ubiquitin, it induced large residual dipolar couplings and pseudocontact shifts that could be measured easily and agreed very well with the protein structure. We show that Cys-Ph-TAHA can be used to label large proteins that are biochemically challenging such as the Lac repressor in a 90 kDa ternary complex with DNA and inducer.

引用

页码：329 / 337

页数：9

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