Redox-Related Chemical Shift Perturbations on Backbone Nuclei of High-Potential Iron−Sulfur Proteins

Lehmann, Teresa; Luchinat, Claudio; Piccioli, Mario

doi:10.1021/ic010761i

Cited by 10 publications

(9 citation statements)

References 47 publications

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“…It should, however, be considered that metal replacement may induce structural changes and consequently chemical shift variations. Also, redox state and charge-dependent chemical shift changes can occur (Vathyam et al 2000 ;Lehmann et al 2002) ; they may affect the diamagnetic term of the observed shift and constitute a bias for the determination of d pc . Indeed, it has been shown that, in Ca 2+ -binding proteins, upon replacement with the diamagnetic La 3+ ion, the resonances of the residues belonging to the metal coordination sphere experience some shift (Allegrozzi et al 2002).…”

Section: Contact Shifts and Relaxation Rates As Restraintsmentioning

confidence: 99%

NMR structures of paramagnetic metalloproteins

Arnesano

Banci

Piccioli

2005

Quart. Rev. Biophys.

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Metalloproteins represent a large share of the proteome and many of them contain paramagnetic metal ions. The knowledge, at atomic resolution, of their structure in solution is important to understand processes in which they are involved, such as electron transfer mechanisms, enzymatic reactions, metal homeostasis and metal trafficking, as well as interactions with their partners. Formerly considered as unfeasible, the first structure in solution by nuclear magnetic resonance (NMR) of a paramagnetic protein was obtained in 1994. Methodological and instrumental advancements pursued over the last decade are such that NMR structure of paramagnetic proteins may be now routinely obtained. We focus here on approaches and problems related to the structure determination of paramagnetic proteins in solution through NMR spectroscopy. After a survey of the background theory, we show how the effects produced by the presence of a paramagnetic metal ion on the NMR parameters, which are in many cases deleterious for the detection of NMR spectra, can be overcome and turned into an additional source of structural restraints. We also briefly address features and perspectives given by the use of 13C-detected protonless NMR spectroscopy for proteins in solution. The structural information obtained through the exploitation of a paramagnetic center are discussed for some Cu2+ -binding proteins and for Ca2+ -binding proteins, where the replacement of a diamagnetic metal ion with suitable paramagnetic metal ions suggests novel approaches to the structural characterization of proteins containing diamagnetic and NMR-silent metal ions.

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Section: Contact Shifts and Relaxation Rates As Restraintsmentioning

confidence: 99%

NMR structures of paramagnetic metalloproteins

Arnesano

Banci

Piccioli

2005

Quart. Rev. Biophys.

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“…In the new millennium, microbiologists, cell and molecular biologists, and eventually geneticists entered into the scenario, affording the study of the pathways for the cluster biosynthesis in Fe-S proteins in model organisms and in humans, thus moving the frontier in Fe-S protein research toward a system-wide perspective (Lill, 2009;Rouault and Tong, 2008;Schmucker and Puccio, 2010). Within this context, where cell biology, integrated structural biology, metalloproteomics, and spectroscopy form a unique research platform that provides a molecular view of Fe-S protein assembly processes and trafficking pathways, paramagnetic NMR contributes to characterizing proteins involved in the Fe-S assembly machineries.…”

Section: Introductionmentioning

confidence: 99%

The long-standing relationship between paramagnetic NMR and iron–sulfur proteins: the mitoNEET example. An old method for new stories or the other way around?

et al. 2021

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Abstract. Paramagnetic NMR spectroscopy and iron–sulfur (Fe–S) proteins have maintained a synergic relationship for decades. Indeed, the hyperfine shifts with their temperature dependencies and the relaxation rates of nuclei of cluster-bound residues have been extensively used as a fingerprint of the type and of the oxidation state of the Fe–S cluster within the protein frame. The identification of NMR signals from residues surrounding the metal cofactor is crucial for understanding the structure–function relationship in Fe–S proteins, but it is generally impaired in standard NMR experiments by paramagnetic relaxation enhancement due to the presence of the paramagnetic cluster(s). On the other hand, the availability of systems of different sizes and stabilities has, over the years, stimulated NMR spectroscopists to exploit iron–sulfur proteins as paradigmatic cases to develop experiments, models, and protocols. Here, the cluster-binding properties of human mitoNEET have been investigated by 1D and 2D 1H diamagnetic and paramagnetic NMR, in its oxidized and reduced states. The NMR spectra of both oxidation states of mitoNEET appeared to be significantly different from those reported for previously investigated [Fe2S2]2+/+ proteins. The protocol we have developed in this work conjugates spectroscopic information arising from “classical” paramagnetic NMR with an extended mapping of the signals of residues around the cluster which can be taken, even before the sequence-specific assignment is accomplished, as a fingerprint of the protein region constituting the functional site of the protein. We show how the combined use of 1D NOE experiments, 13C direct-detected experiments, and double- and triple-resonance experiments tailored using R1- and/or R2-based filters significantly reduces the “blind” sphere of the protein around the paramagnetic cluster. This approach provided a detailed description of the unique electronic properties of mitoNEET, which are responsible for its biological function. Indeed, the NMR properties suggested that the specific electronic structure of the cluster possibly drives the functional properties of different [Fe2S2] proteins.

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“…Since then, many protein structures of paramagnetic systems were obtained in solution in different oxidation states [2][3][4][5][6]. The quest for novel methodological advancements flourished, redox dependent effects were investigated [7], and a number of paramagnetism-based NMR restraints were proposed [8]. Likewise, the main programs for solution structures calculations were revisited and complemented with routines able to tackle paramagnetism-derived NMR restraints and to combine them with conventional NMR restraints [9,10].…”

Section: Introductionmentioning

confidence: 99%

Paramagnetic NMR Spectroscopy Is a Tool to Address Reactivity, Structure, and Protein–Protein Interactions of Metalloproteins: The Case of Iron–Sulfur Proteins

Piccioli¹

2020

Magnetochemistry

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The study of cellular machineries responsible for the iron–sulfur (Fe–S) cluster biogenesis has led to the identification of a large number of proteins, whose importance for life is documented by an increasing number of diseases linked to them. The labile nature of Fe–S clusters and the transient protein–protein interactions, occurring during the various steps of the maturation process, make their structural characterization in solution particularly difficult. Paramagnetic nuclear magnetic resonance (NMR) has been used for decades to characterize chemical composition, magnetic coupling, and the electronic structure of Fe–S clusters in proteins; it represents, therefore, a powerful tool to study the protein–protein interaction networks of proteins involving into iron–sulfur cluster biogenesis. The optimization of the various NMR experiments with respect to the hyperfine interaction will be summarized here in the form of a protocol; recently developed experiments for measuring longitudinal and transverse nuclear relaxation rates in highly paramagnetic systems will be also reviewed. Finally, we will address the use of extrinsic paramagnetic centers covalently bound to diamagnetic proteins, which contributed over the last twenty years to promote the applications of paramagnetic NMR well beyond the structural biology of metalloproteins.

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Redox-Related Chemical Shift Perturbations on Backbone Nuclei of High-Potential Iron−Sulfur Proteins

Cited by 10 publications

References 47 publications

NMR structures of paramagnetic metalloproteins

NMR structures of paramagnetic metalloproteins

The long-standing relationship between paramagnetic NMR and iron–sulfur proteins: the mitoNEET example. An old method for new stories or the other way around?

Paramagnetic NMR Spectroscopy Is a Tool to Address Reactivity, Structure, and Protein–Protein Interactions of Metalloproteins: The Case of Iron–Sulfur Proteins

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