This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
Facts about NADH-ubiquinone oxidoreductase chain 5.
The immediate electron acceptor for the enzyme is believed to be ubiquinone (By similarity). .
Human | |
---|---|
Gene Name: | MT-ND5 |
Uniprot: | P03915 |
Entrez: |
Belongs to: |
---|
complex I subunit 5 family |
NADH-ubiquinone oxidoreductase chain 5
Mass (kDA):
67.027 kDA
Human | |
---|---|
Location: | |
Sequence: | ; |
Mitochondrion inner membrane; Multi-pass membrane protein.
This article will discuss Boster Bio's MT ND5 Marker and Western blotting techniques. Keep reading if Western blotting is new to you or you don't know what the MT-ND5 marker does. In this article, I'll show you the best uses of the MT-ND5 Marker for Western blotting.
MT-ND5 a gene that instructs the production of a NADH dehydrogenase 5 protein. The complex I enzyme compound includes the NADH Dehydrogenase 5 protein. It is active in mitochondria, structures found inside cells that convert energy from food to energy. It is involved in oxidative phosphorylation which uses oxygen, simple sugars, and other chemicals to produce ATP. ATP is the cell’s primary source of energy.
The MT-ND5 gene encodes a protein called NADH dehydrogenase 5. The protein is active in mitochondria which are the structures that make up our cells. They create energy from food by oxidative phosphorylation. This uses oxygen and simple sugars in order to make ATP. ATP is the cell's primary source of energy. The mitochondria produce ATP to increase energy production. This is then converted into glucose.
Four primer pairs were used to amplify the MT ND5 gene. These primer pairs were created using the revised NCBI GenBank Cambridge Reference Sequence by Andrews et al. 1999 data. 100 ng each of the DNA samples were used as a template for the MTND5 marker. Each sample contained 50 uL. We were able amplify this gene successfully using primer pairs which have been shown to reliably produce MTND5 PCR products.
The presence and location of the double peak were checked in the sequencing analysis. BLAST analysis revealed that these two sequences share different nucleotide site. This allowed us to identify genes in the sequencing analysis. However, the MTND5 Gene was not found in either of these samples. This suggests that there is only one nucleotide variation between the two samples for accurate results.
The MTND5 gene is the most abundant in the human genome. The mitochondrial function is responsible for controlling the expression of this marker within cells. This gene can cause a variety of conditions, including mitochondrial diseases, leukemia, renal failure, and other disorders. The gene was first discovered in human embryonic stem cell cells. Heteroplasmic mutations in the MT-ND genes can cause disease. The severity and extent of the disease affects the level of heteroplasmy present in the pathogenic variants.
P8: A patient with a novel mutation was identified in MT–ND5. This patient was previously asymptomatic for 10 years. She suffered frequent migraine attacks, and also had secondary epilepsy. CSF lactate increased to 4.2 mg/l, while CSFmethylation was 2.3 mg/l. Optic nerves appeared pale in ophthalmoscopy. Perimetric examination revealed multiple bilateral Scotomas. Visual evoked Potentials displayed decreased amplitudes and prolonged latency.
The ND5 proteins were isolated from cells and lysed at pH 7.4 in 10 mM Tris/HCl pH 74. After lysis the mitochondrial fraction was stained with a chemiluminescence. On a 10% acrylamide acrylamide chromatogram, the ND5 GFP tagged proteins was found. This protein was then isolated by SDS–PAGE and fluorescent imagery. This protein is found in both mitochondrial muscle and skeletal muscular tissue.
E. coli was used to express ND5 in S. cerevisiae. The amino acid sequence for human ND5 protein was preserved in the synthetic MT–ND5 gene. This gene was created to replace human ND5 protein with a synthetic one. A cloned ND5 gene can be expressed in E. coli and S. cerevisiae.
Patients with multisystem MDs are susceptible to heteroplasmic mtDNA mutations that affect MT-ND5 genes. One patient had a MT-3 mutation while another had MELAS. This mutation may be a sign of fibroblast dysfunction. We have shown that heteroplasmic mutations of the MT-ND5 gene can be associated with the development mitochondrial diseases.
ND5 was copurified with cytochrome c oxidase in mitochondria. We demonstrated that ND5 inserts into the inner mitochondrial membrane. However, the hydrophobicity of the MTS-ND5-Flag fusion protein impeded its mobilization from SDS gels. To immunostain mitochondrial fractions, we used a native MTS–ND5–Flag. S. cerevisiae was not affected by ND5 in its inner mitochondrial membrane.
E145V, a mutant of ND5, interferes with the Li+ detoxification system in S. cerevisiae. Moreover, this mutant mutated in MT-ND5 has a pronounced toxicity. This suggests that MT5's protein plays a significant role in alkali-metal cation transport. The E145V mutant of ND5 also causes less cell growth that the wild type.
PMID: 7219534 by Anderson S., et al. Sequence and organization of the human mitochondrial genome.
PMID: 7530363 by Horai S., et al. Recent African origin of modern humans revealed by complete sequences of hominoid mitochondrial DNAs.