A systematic mutagenesis-driven strategy for site-resolved NMR studies of supramolecular assemblies

Carlos Amero; M. Asuncion Dura; Marjolaine Noirclerc-Savoye; Arnaud Perollier; Benoit Gallet; Michael J. Plevin; Thierry Vernet; Bruno Franzetti; Jerome Boisbouvier

doi:10.1007/s10858-011-9513-5

A systematic mutagenesis-driven strategy for site-resolved NMR studies of supramolecular assemblies

Carlos Amero, M. Asuncion Dura, Marjolaine Noirclerc-Savoye, Arnaud Perollier, Benoit Gallet, Michael J. Plevin, Thierry Vernet, Bruno Franzetti, Jerome Boisbouvier

Biology

Research output: Contribution to journal › Article › peer-review

Abstract

Obtaining sequence-specific assignments remains a major bottleneck in solution NMR investigations of supramolecular structure, dynamics and interactions. Here we demonstrate that resonance assignment of methyl probes in high molecular weight protein assemblies can be efficiently achieved by combining fast NMR experiments, residue-type-specific isotope-labeling and automated site-directed mutagenesis. The utility of this general and straightforward strategy is demonstrated through the characterization of intermolecular interactions involving a 468-kDa multimeric aminopeptidase, PhTET2.

Original language	English
Pages (from-to)	229-236
Number of pages	8
Journal	JOURNAL OF BIOMOLECULAR NMR
Volume	50
Issue number	3
DOIs	https://doi.org/10.1007/s10858-011-9513-5
Publication status	Published - Jul 2011

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abstract = "Obtaining sequence-specific assignments remains a major bottleneck in solution NMR investigations of supramolecular structure, dynamics and interactions. Here we demonstrate that resonance assignment of methyl probes in high molecular weight protein assemblies can be efficiently achieved by combining fast NMR experiments, residue-type-specific isotope-labeling and automated site-directed mutagenesis. The utility of this general and straightforward strategy is demonstrated through the characterization of intermolecular interactions involving a 468-kDa multimeric aminopeptidase, PhTET2.",

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AU - Amero, Carlos

AU - Dura, M. Asuncion

AU - Noirclerc-Savoye, Marjolaine

AU - Perollier, Arnaud

AU - Gallet, Benoit

AU - Plevin, Michael J.

AU - Vernet, Thierry

AU - Franzetti, Bruno

AU - Boisbouvier, Jerome

PY - 2011/7

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AB - Obtaining sequence-specific assignments remains a major bottleneck in solution NMR investigations of supramolecular structure, dynamics and interactions. Here we demonstrate that resonance assignment of methyl probes in high molecular weight protein assemblies can be efficiently achieved by combining fast NMR experiments, residue-type-specific isotope-labeling and automated site-directed mutagenesis. The utility of this general and straightforward strategy is demonstrated through the characterization of intermolecular interactions involving a 468-kDa multimeric aminopeptidase, PhTET2.

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JF - JOURNAL OF BIOMOLECULAR NMR

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ER -

A systematic mutagenesis-driven strategy for site-resolved NMR studies of supramolecular assemblies

Abstract

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