Recombinant Inia geoffrensis NADH-ubiquinone oxidoreductase chain 4L (MT-ND4L)
Description
Introduction to MT-ND4L
NADH-ubiquinone oxidoreductase chain 4L (MT-ND4L) is a core subunit of Complex I, a proton-pumping enzyme in the mitochondrial electron transport chain (ETC) responsible for transferring electrons from NADH to ubiquinone. This process generates a proton gradient critical for ATP synthesis . In Inia geoffrensis (a freshwater dolphin), recombinant MT-ND4L is produced via bacterial expression systems (e.g., E. coli) to study its structure, function, and potential biomedical applications .
2.1. Protein Properties
2.2. Role in Complex I
MT-ND4L is part of the transmembrane domain of Complex I, which:
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Transfers electrons from NADH to ubiquinone via FMN and Fe-S clusters.
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Pumps protons across the mitochondrial membrane, contributing to the proton gradient .
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Interacts with other subunits (e.g., ND1, ND2, ND3, ND4, ND5, ND6) to form the enzyme’s active site .
3.1. Production and Purification
Recombinant MT-ND4L from Inia geoffrensis is synthesized in E. coli and purified using:
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Chromatography: IMAC (Immobilized Metal Affinity Chromatography) for His-tagged proteins .
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Tags: N-terminal His6-ABP or other fusion tags to enhance solubility and stability .
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Purity: >80% as confirmed by SDS-PAGE and Coomassie blue staining .
3.2. Applications in Studies
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Antibody Validation: Used as a blocking antigen to confirm antibody specificity in immunoassays .
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Structural Analysis: Amino acid sequences (e.g., LLVSISNTYGLDYVHNLNLLQ) aid in modeling hydrophobic interactions within Complex I .
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Electron Transfer Studies: Mutagenesis and biochemical assays to elucidate ubiquinone binding and proton translocation .
4.1. Functional Insights
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Electron Transfer: MT-ND4L’s hydrophobic residues stabilize the enzyme’s membrane domain, enabling efficient electron relay .
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Proton Pumping: Mutations in MT-ND4L (e.g., Val65Ala in humans) disrupt proton translocation, linking to mitochondrial diseases like Leber’s Hereditary Optic Neuropathy (LHON) .
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Species-Specific Variations: Inia geoffrensis MT-ND4L shares ~60–70% sequence identity with human ND4L, suggesting conserved core functions but divergent regulatory regions .
4.2. Limitations and Knowledge Gaps
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Sparse Data: Limited studies on Inia geoffrensis MT-ND4L; most research focuses on human, Chlamydomonas, or bacterial homologs .
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Structural Complexity: Full-length recombinant MT-ND4L is challenging to produce due to its hydrophobic nature, necessitating strategic solubility tags .
Clinical and Biomedical Relevance
Product Specs
Please note that we will prioritize shipping the format currently in stock. However, if you have specific requirements for the format, kindly indicate them in your order remarks, and we will accommodate your request.
Our proteins are shipped with standard blue ice packs by default. If you require dry ice shipping, please communicate with us in advance, as additional fees may apply.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. For lyophilized form, the shelf life is 12 months at -20°C/-80°C.
