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25 Jan 2023

A novel anti-Nduf11a antibody provides vital insight

University of Bristol

Mitochondrial dysregulation can have devastating and fatal pathologies. Research aims to understand the function of respiratory complexes to bring a brighter future to those afflicted by these conditions. Significant progress has been made through developing a highly-specific antibody anti-Nduf11a by Cambridge Research Biochemicals for Professor Ian Collinson to perform mitochondrial respiratory complex analysis.

Mitochondria provide energy for the cell in the form of ATP via the electron transport chain step-wise transfer through four large multimeric electron transfer complexes (CI-IV). The complexes form higher-order assemblies with specific stoichiometry. Several supercomplexes have been detected showing a conserved core of monomeric CI and dimeric CIII (CI1:III2). With strong conservation, it implies this core complex has a vital role in mitochondria but it has yet to be understood.

At the interface of the CI1:III2 complex is an integral membrane protein, named NDUFA11 in mammals. NDUFA11 is believed to provide a critical interaction within the supercomplex and complex I assembly. Mutated NDUFA11 results in CI deficiency that leads to several pathologies including fatal infantile lactic acidemia, encephalocardiomyopathy, and late-onset myopathy. The importance of NDUFA11 in the higher-order stability of supercomplexes has not been established.

Professor Ian Collinson and his team at the University of Bristol decided to explore the CI1:III2 complex by using the genetically tractable model organism, Caenorhabditis elegans. The team successfully confirmed the presence of the C. elegans NDUFA11 homologue and named it NDUF-11. C. elegans, nduf-11 knockout arrests at an early larval stage. Therefore, the next step was to try NDUF-11 depletion using RNAi, this should allow the organism to reach adulthood to try and understand the reason for the lethality.

Ian Collinson contacted Cambridge Research Biochemicals to produce a custom antibody for this crucial target, NDUF-11. “CRB were friendly and approachable while providing technical and design advice at every step of antibody production. The after-support and communication were greatly appreciated and the antibody was incredibly useful.”

The highly specific anti-Nduf11a antibody can detect NDUF-11 within the CI complex while demonstrating that RNAi treatment can deplete the levels. The anti-Nduf11a antibody provided vital evidence that NDUF-11 is integral to the mitochondrial CI complex, and significantly, reduced levels of the NDUF-11 result in destabilised CI and supercomplexes, leading to the deregulation of respiration. 

Depletion of NDUF-11 levels with RNAi. Anti-Nduf11a antibody successfully detects Nduf11 and the significant reduction of Nduf11 protein in mitochondria after RNAi treatment.
Depletion of NDUF-11 levels with RNAi. Anti-Nduf11a antibody successfully detects Nduf11 and the significant reduction of Nduf11 protein in mitochondria after RNAi treatment. 50 µg of isolated mitochondria were loaded per lane, ATP-5 was used as a loading control. **P<0.01 (Student’s t-test).

Ian Collinson’s team used multiple methods of investigation and had the resource of the custom-made specific antibody anti-Nduf11a. Together this provided evidence of the requirement of NDUF-11 for normal respiratory function, CI assembly, and supercomplex maintenance. The highly specific anti-Nduf11a antibody helps the understanding of the underlying mechanisms that are perturbed in mitochondrial disease by a failure of respiratory complex integrity.

Ian Collinson had this to say, “working with CRB on this antibody gave me the confidence to do so again on another anti-peptide antibody project. I highly recommend them.”

Citation: Knapp-Wilson et al., (2021). Maintenance of complex I and its supercomplexes by NDUF-11 is essential for mitochondrial structure, function and health. J. Cell Sci., 134(13): jcs258399. PMID: 34106255.