Exploring the FMN binding site in the mitochondrial outer membrane protein mitoNEET

Tasnim, Homyra; Landry, Aaron P.; Fontenot, Chelsey R.; Ding, Huangen

doi:10.1016/j.freeradbiomed.2020.05.004

Cited by 13 publications

(13 citation statements)

References 55 publications

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“…Two-dimensional standard NMR and circular dichroism (CD) spectra acquired in the mitoNEET C-terminal cytosolic domain (res 44-108) showed that the unique fold adopted by each subunit in the holo protein is strictly related to the presence of a cluster that can be disassembled and reassembled in vitro, inducing, respectively, unfolding and refolding of the protein (Ferecatu et al, 2014). EPR spectroscopy, performed on E. coli cells containing the overexpressed cytosolic domain of human mitoNEET, showed that in the cytoplasmic cellular environment the two [Fe 2 S 2 ] clusters are in the reduced state (Landry and Ding, 2014), as expected given their ∼ 0 mV midpoint redox potential, measured in vitro at pH 7.5 (Bak et al, 2009;Tirrell et al, 2009). The oxidation state of the [Fe 2 S 2 ] clusters of mitoNEET plays a crucial role in the in vitro cluster transfer activity of the protein, since only [Fe 2 S 2 ] 2+ and not [Fe 2 S 2 ] + cluster transfer from holo mi-toNEET to apo recipient proteins has been observed (Ferecatu et al, 2014;Lipper et al, 2015;Zuris et al, 2011).…”

Section: Mitoneet: (Another) Protein In Search Of a Function?supporting

confidence: 57%

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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Section: Mitoneet: (Another) Protein In Search Of a Function?supporting

confidence: 57%

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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“…This was consistent with the overall mitochondrial function and cardiac function in the 3-month-old mice. Furthermore, it has been known that mitoNEET functions as electron-transfer protein 9,[25][26][27] . The reduced flavin mononucleotide interacts with mitoNEET via specific binding site and transfers its electrons to the 2Fe-2S clusters of mitoNEET, and the reduced 2Fe-2S clusters in mito-NEET transfer the electrons to oxygen or ubiquinone.…”

Section: Discussionmentioning

confidence: 99%

Cardiac-specific loss of mitoNEET expression is linked with age-related heart failure

et al. 2021

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Heart failure (HF) occurs frequently among older individuals, and dysfunction of cardiac mitochondria is often observed. We here show the cardiac-specific downregulation of a certain mitochondrial component during the chronological aging of mice, which is detrimental to the heart. MitoNEET is a mitochondrial outer membrane protein, encoded by CDGSH iron sulfur domain 1 (CISD1). Expression of mitoNEET was specifically downregulated in the heart and kidney of chronologically aged mice. Mice with a constitutive cardiac-specific deletion of CISD1 on the C57BL/6J background showed cardiac dysfunction only after 12 months of age and developed HF after 16 months; whereas irregular morphology and higher levels of reactive oxygen species in their cardiac mitochondria were observed at earlier time points. Our results suggest a possible mechanism by which cardiac mitochondria may gradually lose their integrity during natural aging, and shed light on an uncharted molecular basis closely related to age-associated HF.

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“…Actually, Rieske proteins and the plant type ferredoxins (which share the same NMR features) act as electron transfer proteins, while ISCA1 and ISCA2 are involved into the assembly and transfer of the cluster. MitoNEET is supposed to play a major role in restoring the Fe-S cluster on cytosolic apo aconitase IRP1 in oxidative stress conditions (Ferecatu et al, 2014), and acts as a cluster transfer protein for several apo recipient proteins (Ferecatu et al, 2014;Lipper et al, 2015;Zuris et al, 2011), both functions being based on a redox switch, activated by several cellular cofactors (Camponeschi et al, 2017;Tasnim et al, 2020;Wang et al, 2017). These findings are intriguing: a different coordination structure of the cluster, which determines the valence localization/delocalization within the cluster may be the origin of its different electronic properties, thus determining different redox partner.…”

Section: Discussionmentioning

confidence: 99%

“…However, it has been proposed that, for most of the cellular functions, the [Fe2S2] clusters of dimeric mitoNEET might play a crucial role, possibly acting as redox-or pH-sensors for mitochondrial functions, and/or being transferred to cytosolic apo proteins in response to the redox states of the cells (Ferecatu et al, 2014;Lipper et al, 2015;Zuris et al, 2011). Indeed, mitoNEET shows redox activity in vitro (Camponeschi et al, 2017;Landry and Ding, 2014;Tasnim et al, 2020;Wang et al, 2017), and is able to repair Fe-S proteins, by reloading Fe-S clusters onto cytosolic proteins whose Fe-S clusters have been removed or altered (Ferecatu et al, 2014). The electronic properties and chemical reactivity of mitoNEET clusters have been extensively investigated so far through several biophysical and biochemical techniques.…”

Section: Mitoneet: (Another) Protein In Search Of a Function?mentioning

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?

Camponeschi¹,

Gallo²,

Piccioli³

et al. 2021

Preprint

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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 size and stability 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 one-dimensional and two-dimensional 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. We show how the use of 1D NOE experiments, 13C direct-detected experiments, and the optimization of NMR experiments for paramagnetic systems significantly reduce the blind sphere of the protein around the paramagnetic cluster. The application of this approach provided a detailed description of the unique electronic properties of mitoNEET, that 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.

show abstract

Exploring the FMN binding site in the mitochondrial outer membrane protein mitoNEET

Cited by 13 publications

References 55 publications

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?

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?

Cardiac-specific loss of mitoNEET expression is linked with age-related heart failure

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?

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