TDP-43, FUS, SOD1 Localization and Aggregation Assay Service

At the heart of amyotrophic lateral sclerosis (ALS) pathology lies the mislocalization and aggregation of critical proteins: TDP-43, FUS, and SOD1. Our services leverage globally recognized, cutting-edge technologies and a comprehensive, multi-dimensional analytical approach. We empower researchers to dissect these pathological protein behaviors, accelerating the discovery of novel drug targets and the development of robust diagnostic markers. For further information regarding the products and services provided, project-specific consultation, and pricing, please submit an inquiry here.

Aggregation-Prone Proteins in ALS

• TDP-43 and FUS: Key Players in RNA Metabolism Regulation

The precise choreography of TDP-43 and FUS between the nucleus and cytoplasm, governed by their respective nuclear localization signals (NLS) and nuclear export signals (NES), is critical for cellular homeostasis. Notably, the glycine-rich region of TDP-43 and the SYGQ domain of FUS are instrumental in their RNA binding and transport activities.

In ALS, TDP-43 forms ubiquitinated and phosphorylated aggregates, with the C-terminal fragment, such as TDP-25, exhibiting significant neurotoxicity. Conversely, mutations within FUS disrupt nucleocytoplasmic transport, driving its aberrant cytoplasmic accumulation and aggregation. These disruptions highlight the delicate balance of nucleocytoplasmic trafficking and the profound consequences when this balance is compromised in neurodegenerative diseases.

Fig.1 Structures and functional domains of TDP-43 and FUS proteins.^{1, 4}

• SOD1: A Core Driver of Familial ALS

Mutations within the copper/zinc catalytic active site of superoxide dismutase 1 (SOD1) trigger misfolding, culminating in the formation of amyloid aggregates and direct neuronal damage. Notably, SOD1 mutants, such as G93A, induce motor neuron apoptosis through the formation of insoluble aggregates, with their toxicity manifesting independently of wild-type SOD1. Furthermore, the metal binding state of SOD1, particularly demetallation, exerts a substantial influence on its aggregation propensity.

Fig.2 Structures and functional domains of SOD1 protein.^{1, 4}

• Multi-Protein Coaggregation

The synergistic cytoplasmic co-aggregation of TDP-43, FUS, and SOD1 disrupts the endoplasmic reticulum-neurofilament transport system, culminating in synaptic function loss. Given that all three proteins participate in miRNA metabolism, detecting their aberrant localization offers a powerful approach to elucidating the molecular mechanisms underlying RNA homeostasis imbalance in ALS.

Fig.3 SOD1 Misfolding in SOD1-FALS, FUS-FALS and SALS with TDP43 Pathology.^{2, 4}

Service Features and Technology Highlights

• Comprehensive, End-to-End Solutions

Precise Localization Analysis: Utilizing highly specific antibodies coupled with advanced fluorescent labeling, we deliver high-resolution visualization of subcellular protein localization.

Accurate Aggregation Quantification: We employ a multi-faceted approach, combining thioflavin T (THT) fluorescence staining, protein blotting, and ELISA, to precisely determine the ratio of soluble to insoluble proteins, providing critical insights into aggregation dynamics.

Real-Time Dynamic Tracking: Through live cell imaging, we monitor the dynamic processes of aggregate formation and propagation, allowing for a deeper understanding of disease progression.

Fig.4 Aggregated mutant TDP-43 and FUS.^{3, 4}

• High-Throughput Capabilities

Human Pluripotent Stem Cell (hiPSC)-Derived Motor Neuron Models: We have established robust culture systems using motor neurons derived from human pluripotent stem cells, both from control and patient lines. This enables efficient, high-throughput screening for potential therapeutic targets and disease modifiers.

Our technical team has many years of experience in neurodegenerative disease research and is committed to providing highly sensitive and reproducible detection solutions for global research institutions and pharmaceutical companies. Please contact us for a customized experimental design!

References

1. Duranti, Elisa, and Chiara Villa. "Molecular investigations of protein aggregation in the pathogenesis of amyotrophic lateral sclerosis." International journal of molecular sciences 24.1 (2022): 704.
2. Pokrishevsky, Edward, et al. "Aberrant localization of FUS and TDP43 is associated with misfolding of SOD1 in amyotrophic lateral sclerosis." PloS one 7.4 (2012): e35050.
3. Farrawell, Natalie E., et al. "Distinct partitioning of ALS associated TDP-43, FUS and SOD1 mutants into cellular inclusions." Scientific reports 5.1 (2015): 13416.
4. Distributed under Open Access license CC BY 4.0, without modification.