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Combined Biochemical, Biophysical, and Cellular Methods to Study Fe-S Cluster Transfer and Cytosolic Aconitase Repair by MitoNEET.

Methods Enzymol.. 2017; 
Mons C, Ferecatu I, Riquier S, Lescop E, Bouton C, Golinelli-Cohen MP.
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PCR Cloning and Subcloning ... Paddock et al., 2007). Codon usage was optimized for its expression in E. coli (GenScript). The open reading frame coding for the fragment M44-T108 was cloned between NdeI and XhoI of pET22b(+) vector. Using this construct ... Get A Quote
Bacterial Expression System Get A Quote

摘要

MitoNEET is the first identified Fe-S protein anchored to mammalian outer mitochondrial membranes with the vast majority of the protein polypeptide located in the cytosol, including its [2Fe-2S] cluster-binding domain. The coordination of the cluster is unusual and involves three cysteines and one histidine. MitoNEET is capable of transferring its redox-active Fe-S cluster to a bacterial apo-ferredoxin in vitro even under aerobic conditions, unlike other Fe-S transfer proteins such as ISCU. This specificity suggests its possible involvement in Fe-S repair after oxidative and/or nitrosative stress. Recently, we identified cytosolic aconitase/iron regulatory protein 1 (IRP1) as the first physiological protein acc... More

关键词

Cluster disassembly; Cluster repair; Cluster transfer; Cytosolic aconitase; MitoNEET; Nuclear magnetic resonance spectroscopy; UV–visible absorption spectroscopy; siRNA