NDUFS5 Human
Description
NDUFS5 is fused to a 23 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.
Product Specs
Product Science Overview
Histidine NADH Dehydrogenase Fe-S Protein 5, also known as NDUFS5, is a subunit of the NADH dehydrogenase (ubiquinone) complex. This complex plays a crucial role in cellular respiration by transferring electrons from NADH to ubiquinone, a process that contributes to the generation of ATP through oxidative phosphorylation. The human recombinant form of this protein is produced using recombinant DNA technology, which allows for the expression of the protein in a host organism, typically bacteria or yeast.
NDUFS5 is an iron-sulfur (Fe-S) protein, which means it contains iron-sulfur clusters that are essential for its electron transfer function. These clusters facilitate the transfer of electrons within the complex, contributing to the overall efficiency of the electron transport chain. The histidine tag in the recombinant form of the protein aids in its purification and detection.
The preparation of human recombinant NDUFS5 involves several steps:
- Gene Cloning: The gene encoding NDUFS5 is cloned into an expression vector, which is then introduced into a host organism.
- Protein Expression: The host organism, often Escherichia coli, is cultured under conditions that induce the expression of the recombinant protein.
- Purification: The histidine tag allows for the purification of the protein using affinity chromatography, where the protein binds to a column containing nickel or cobalt ions.
- Verification: The purity and functionality of the protein are verified using techniques such as SDS-PAGE and Western blotting.
NDUFS5 participates in the redox reactions of the electron transport chain. The iron-sulfur clusters within the protein undergo oxidation and reduction as they transfer electrons from NADH to ubiquinone. This process is coupled with the translocation of protons across the mitochondrial membrane, contributing to the proton gradient that drives ATP synthesis.
The activity of NDUFS5 can be analyzed using spectrophotometric assays that measure the reduction of ubiquinone or the oxidation of NADH. Additionally, the integrity of the iron-sulfur clusters can be assessed using electron paramagnetic resonance (EPR) spectroscopy.
Recombinant NDUFS5 is used in various research applications, including:
- Studying Mitochondrial Function: Understanding the role of Complex I in cellular respiration and its implications in mitochondrial diseases.
- Drug Development: Screening for compounds that can modulate the activity of Complex I, which may have therapeutic potential for conditions such as neurodegenerative diseases and cancer.
- Biochemical Research: Investigating the structure-function relationships of iron-sulfur proteins and their role in electron transfer processes.
