NDUFB7 Antibody

Introduction to NDUFB7 Protein and Its Antibodies

NDUFB7, also known as Complex I-B18 or NADH-ubiquinone oxidoreductase B18 subunit, is a nuclear-encoded accessory subunit of the mitochondrial electron transport chain Complex I. This 16 kDa protein plays a crucial role in cellular respiration and energy production . NDUFB7 is characterized by:

• A calculated molecular weight of 16 kDa, though often observed at 18-22 kDa in electrophoretic analyses

• Localization to the mitochondrial inner membrane

• A sevenfold repeat of positively charged residues suggesting involvement in protein-protein interactions

• A sequence of 137 amino acids in humans

• A unique N-myristoylation motif that is highly conserved among vertebrates

NDUFB7 antibodies are immunological reagents specifically designed to detect this protein in various experimental applications. These antibodies have become essential tools in mitochondrial research, enabling scientists to investigate the structure, function, and pathological relevance of NDUFB7 and Complex I.

Types and Properties of NDUFB7 Antibodies

NDUFB7 antibodies are available in various formats to suit different research applications. The key properties of commercially available NDUFB7 antibodies are summarized in the following table:

The diversity of available NDUFB7 antibodies allows researchers to select the most appropriate reagent for their specific experimental needs, contributing to the advancement of mitochondrial research.

Applications of NDUFB7 Antibodies in Research

NDUFB7 antibodies are utilized in multiple experimental techniques to study mitochondrial structure and function. The major applications include:

Western Blot Analysis

Western blotting is one of the most common applications for NDUFB7 antibodies. The recommended dilutions typically range from 1:500 to 1:50,000, depending on the specific antibody and sample type . Western blot analysis with NDUFB7 antibodies has been used to:

• Detect NDUFB7 protein expression in various cell lines and tissues

• Confirm the molecular weight of NDUFB7 (observed at 16-18 kDa)

• Examine alterations in NDUFB7 expression under different experimental conditions

• Investigate Complex I assembly in patients with mitochondrial disorders

Immunohistochemistry

NDUFB7 antibodies are effectively used in immunohistochemistry to visualize the distribution and expression of NDUFB7 in tissue sections. The recommended dilutions typically range from 1:20 to 1:1,000 . Applications include:

• Examination of NDUFB7 expression in normal and diseased tissues

• Analysis of mitochondrial distribution in different cell types

• Visualization of NDUFB7 in paraffin-embedded tissue samples from human, mouse, and rat origins

Immunofluorescence

Immunofluorescence is utilized to determine the subcellular localization of NDUFB7. This technique has been particularly valuable in research examining:

• Co-localization of NDUFB7 with other mitochondrial proteins

• Alterations in mitochondrial morphology and distribution

• The impact of NDUFB7 mutations on protein localization

• The role of N-myristoylation in NDUFB7 targeting to mitochondria

Additional Applications

NDUFB7 antibodies have also been employed in several other techniques:

• Immunoprecipitation (IP) to isolate NDUFB7 and its interacting proteins

• Flow cytometry to analyze NDUFB7 expression in cell populations

• ELISA for quantitative determination of NDUFB7 levels

• Blue Native PAGE to study Complex I assembly and supercomplexes

Mitochondrial Complex Assembly Studies

Research using NDUFB7 antibodies has revealed crucial insights into mitochondrial Complex I assembly and function. A significant finding from these studies is the demonstration that fibroblasts from patients with NDUFB7 mutations show deficient Complex I assembly and reduced supercomplex formation . Blue native PAGE analysis revealed that:

• Complex I is not visible in patient samples using strong detergents like Triton X-100

• Complexes III and IV amounts are also reduced in affected patients

• Supercomplexes I, III, and IV are significantly reduced in patients with NDUFB7 mutations

• Two-dimensional BN-PAGE demonstrated a profound decrease in Complex I subunit levels

These findings highlight the essential role of NDUFB7 in maintaining the structural integrity and functional capacity of mitochondrial Complex I.

Protein N-myristoylation and Mitochondrial Targeting

A groundbreaking study using NDUFB7 antibodies identified NDUFB7 as one of only four N-myristoylated proteins that specifically localize to mitochondria . This research demonstrated that:

• NDUFB7 undergoes N-myristoylation, a post-translational modification that attaches a myristoyl group to the N-terminal glycine

• Protein N-myristoylation is critical for the mitochondrial localization of NDUFB7

• The CHCH domain in NDUFB7 is essential for its mitochondrial localization

• Non-myristoylatable G2A mutants of NDUFB7 localize exclusively to the nucleus instead of mitochondria

These findings have shed light on the complex mechanisms governing protein targeting to mitochondria and have implications for understanding mitochondrial disorders associated with NDUFB7 dysfunction.

Disease-Associated Research

NDUFB7 antibodies have been instrumental in studies investigating the pathological consequences of NDUFB7 mutations. Recent research has identified patients with compound heterozygous mutations in NDUFB7 who present with:

• Brain neuronal defects

• Lactic acidosis

• Pons abnormality

• Prenatal and postnatal growth deficiency

• Incomplete closure of the abdominal wall

• Poorly functioning gastrointestinal tract

In vivo studies using zebrafish models with knocked-down Ndufb7 expression have demonstrated:

• Brain ventricle and neuronal defects

• Elevated lactic acid levels

• Reduced oxygen consumption

• Defective mitochondrial respiration

• Response to MitoQ treatment, suggesting potential therapeutic approaches

NDUFB7-Related Disorders

Research using NDUFB7 antibodies has contributed significantly to our understanding of mitochondrial disorders associated with NDUFB7 dysfunction. Patients with NDUFB7 mutations present with a spectrum of clinical manifestations, including:

• Severe congenital lactic acidosis

• Hypertrophic cardiomyopathy

• Neurological abnormalities

• Growth deficiencies

• Gastrointestinal dysfunction

Therapeutic Approaches

Studies utilizing NDUFB7 antibodies have identified potential therapeutic strategies for NDUFB7-related disorders. Treatment approaches that have shown promise include:

• Coenzyme Q10 supplementation

• Vitamin B complex administration

• Mitoquinone mesylate (MitoQ) therapy, which has demonstrated efficacy in ameliorating phenotypes in zebrafish models of NDUFB7 deficiency

These findings suggest that targeting mitochondrial function may represent a viable therapeutic approach for patients with NDUFB7 mutations and related mitochondrial disorders.

Future Perspectives in NDUFB7 Research

The continued development and application of NDUFB7 antibodies are expected to facilitate several emerging areas of research:

• Further elucidation of the role of protein N-myristoylation in NDUFB7 function and mitochondrial targeting

• Expanded understanding of the protein-protein interactions mediated by NDUFB7 within Complex I

• Development of more effective therapeutic strategies for NDUFB7-related disorders

• Utilization of animal models for drug screening and preclinical testing

• Investigation of the potential role of NDUFB7 in more common neurodegenerative and metabolic disorders

As research tools continue to evolve, NDUFB7 antibodies will remain indispensable for advancing our understanding of mitochondrial biology and pathology.