Optimized co-solute paramagnetic relaxation enhancement for the rapid NMR analysis of a highly fibrillogenic peptide

被引：0

作者：

Nur Alia Oktaviani

Michael W. Risør

Young-Ho Lee

Rik P. Megens

Djurre H. de Jong

Renee Otten

Ruud M. Scheek

Jan J. Enghild

Niels Chr. Nielsen

Takahisa Ikegami

Frans A. A. Mulder

机构：

[1] University of Groningen,Groningen Biomolecular Sciences and Biotechnology Institute

[2] University of Aarhus,Interdisciplinary Nanoscience Center (iNANO) and Department of Chemistry

[3] Osaka University,Institute for Protein Research

[4] University of Groningen,Stratingh Institute for Chemistry

[5] University of Aarhus,Interdisciplinary Nanoscience Center (iNANO) and Department of Molecular Biology and Genetics

[6] Yokohama City University,Graduate School of Medical Life Science

来源：

Journal of Biomolecular NMR | 2015年 / 62卷

关键词：

Paramagnetic relaxation enhancement; Intrinsically disordered proteins; Fe(DO3A); α; -Antitrypsin; Amyloid beta-peptides; NMR spectroscopy;

D O I：

暂无

中图分类号：

学科分类号：

摘要：

Co-solute paramagnetic relaxation enhancement (PRE) is an attractive way to speed up data acquisition in NMR spectroscopy by shortening the T1 relaxation time of the nucleus of interest and thus the necessary recycle delay. Here, we present the rationale to utilize high-spin iron(III) as the optimal transition metal for this purpose and characterize the properties of its neutral chelate form Fe(DO3A) as a suitable PRE agent. Fe(DO3A) effectively reduces the T1 values across the entire sequence of the intrinsically disordered protein α-synuclein with negligible impact on line width. The agent is better suited than currently used alternatives, shows no specific interaction with the polypeptide chain and, due to its high relaxivity, is effective at low concentrations and in ‘proton-less’ NMR experiments. By using Fe(DO3A) we were able to complete the backbone resonance assignment of a highly fibrillogenic peptide from α1-antitrypsin by acquiring the necessary suite of multidimensional NMR datasets in 3 h.

引用

页码：129 / 142

页数：13

共 135 条

[1] Bermel W(2006)13C-detected protonless NMR spectroscopy of proteins in solution Prog Nucl Magn Reson Spectrosc 48 25-45
[2] Bertini I(2006)Protonless NMR experiments for sequence-specific assignment of backbone nuclei in unfolded proteins J Am Chem Soc 128 3918-3919
[3] Felli I(2006)Sensitivity enhancement of multidimensional NMR experiments by paramagnetic relaxation effects J Am Chem Soc 128 13474-13478
[4] Piccioli M(2009)Protein-targeted gadolinium-based magnetic resonance imaging (MRI) contrast agents: design and mechanism of action Acc Chem Res 42 851-862
[5] Pierattelli R(1993)Synthesis, characterization, and crystal structure of M(DO3A) (M = Fe, Gd) and Na[M(DOTA)] (M = Fe, Y, Gd) Inorg Chem 32 3501-3508
[6] Bermel W(1995)NMRPipe: a multidimensional spectral processing system based on UNIX pipes J Biomol NMR 6 277-293
[7] Bertini I(2000)The carboxyl-terminal fragment of alpha1-antitrypsin is present in atherosclerotic plaques and regulates inflammatory transcription factors in primary human monocytes Mol Cell Biol Res Commun 4 50-61
[8] Felli IC(2005)Extended flip-back schemes for sensitivity enhancement in multidimensional HSQC-type out-and-back experiments J Biomol NMR 33 243-259
[9] Lee YM(2000)Topography of a 2.0 Å structure of alpha1-antitrypsin reveals targets for rational drug design to prevent conformational disease Protein Sci 9 1274-1281
[10] Luchinat C(2010)Kinetics of amyloid beta monomer-to-oligomer exchange by NMR relaxation J Am Chem Soc 132 9948-9951

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