The NMR signature of gluconoylation: a frequent N-terminal modification of isotope-labeled proteins

被引：0
作者：
David Schweida
Pierre Barraud
Christof Regl
Fionna E. Loughlin
Christian G. Huber
Chiara Cabrele
Mario Schubert
机构：
[1] University of Salzburg,Department of Biosciences
[2] ETH Zürich,Institute of Molecular Biology and Biophysics
[3] Institut de Biologie Physico-Chimique (IBPC),Christian Doppler Laboratory for Innovative Tools for Biosimilar Characterization
[4] UMR 8261 CNRS,Department of Biochemistry & Molecular Biology, Monash Biomedicine Discovery Institute
[5] Université Paris Diderot,undefined
[6] University of Salzburg,undefined
[7] Monash University,undefined
来源：
Journal of Biomolecular NMR | 2019年 / 73卷
关键词：
NMR spectroscopy; Post-translational protein modification; Gluconoylation; Gluconic acid; N-terminus; Recombinant protein;
D O I：
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中图分类号：
学科分类号：
摘要：
N-terminal gluconoylation is a moderately widespread modification in recombinant proteins expressed in Escherichia coli, in particular in proteins bearing an N-terminal histidine-tag. This post-translational modification has been investigated mainly by mass spectrometry. Although its NMR signals must have been observed earlier in spectra of 13C/15N labeled proteins, their chemical shifts were not yet reported. Here we present the complete 1H and 13C chemical shift assignment of the N-terminal gluconoyl post-translational modification, based on a selection of His-tagged protein constructs (CCL2, hnRNP A1 and Lin28) starting with Met-Gly-...-(His)6. In addition, we show that the modification can hydrolyze over time, resulting in a free N-terminus and gluconate. This leads to the disappearance of the gluconoyl signals and the appearance of gluconate signals during the NMR measurements. The chemical shifts presented here can now be used as a reference for the identification of gluconoylation in recombinant proteins, in particular when isotopically labeled.
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页码：71 / 79
页数：8
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