Glycine specifically labelled proteins to enhance solution NMR of methylene groups
6 affiliations
- 1Laboratoire Chimie Physique et Chimie du Vivant, CPCV
- 2Sorbonne Université, École Normale Supérieure, PSL Université, CNRS
- 3Integrated Structural Biology Grenoble (ISBG), Cell-free Platform
- 4Univ. Grenoble Alpes, CNRS, CEA, EMBL
- 5Institut de Biologie Structurale (IBS)
- 6Université Grenoble Alpes, CNRS, CEA
Abstract
Specific isotopic labelling of proteins has greatly advanced biomolecular NMR. Protein domains, individual residues or certain chemical groups with specific patterns of isotopes can be incorporated to optimize the use of NMR pulse schemes. Here, we describe a novel cell-free protein expression protocol that allows specific isotopic labelling of glycine residues, whilst all other amino acids are independently labelled. This technique enables us to prevent amino acid scrambling and cross-protonation from the solvent resulting in high labelling selectivity of glycine residues as attested by quantification of 15N amide scrambling and residual Ca protonation. We then highlight the potential applications of these samples in the field of biomolecular NMR. We produced a 1 H, 13 C glycine specifically labelled sample of a subunit of the heavy metal ATPase HMA8, in a completely perdeuterated 2 H background and demonstrate how the CH 2 -TROSY experiment recorded on such a sample improves the spectral quality of glycine residues.
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BibTeX
@misc{capello2026glycine,
title = {{Glycine specifically labelled proteins to enhance solution NMR of methylene groups}},
author = {Rachel Lenoir Capello and Lionel Imbert and Jérôme Boisbouvier and Emeric Miclet},
howpublished = {ChemRxiv},
year = {2026},
month = oct,
doi = {10.26434/chemrxiv.15009868/v1},
url = {https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009868/v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Glycine specifically labelled proteins to enhance solution NMR of methylene groups
AU - Rachel Lenoir Capello
AU - Lionel Imbert
AU - Jérôme Boisbouvier
AU - Emeric Miclet
PY - 2026
DA - 2026/10/05
PB - ChemRxiv
DO - 10.26434/chemrxiv.15009868/v1
UR - https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009868/v1
AB - Specific isotopic labelling of proteins has greatly advanced biomolecular NMR. Protein domains, individual residues or certain chemical groups with specific patterns of isotopes can be incorporated to optimize the use of NMR pulse schemes. Here, we describe a novel cell-free protein expression protocol that allows specific isotopic labelling of glycine residues, whilst all other amino acids are independently labelled. This technique enables us to prevent amino acid scrambling and cross-protonation from the solvent resulting in high labelling selectivity of glycine residues as attested by quantification of 15N amide scrambling and residual Ca protonation. We then highlight the potential applications of these samples in the field of biomolecular NMR. We produced a 1 H, 13 C glycine specifically labelled sample of a subunit of the heavy metal ATPase HMA8, in a completely perdeuterated 2 H background and demonstrate how the CH 2 -TROSY experiment recorded on such a sample improves the spectral quality of glycine residues.
ER -