KCC2 (K+-Cl- Cotransporter 2) is critical for several essential neuronal processes. It plays a pivotal role in the extrusion of chloride ions from neurons, which is vital for maintaining the proper electrochemical gradient necessary for neurotransmission. This function is particularly important in shaping cortical development and regulating GABAergic synaptic strength, which influences neuronal plasticity and network functions. By controlling chloride levels, KCC2 helps ensure that GABA, the primary inhibitory neurotransmitter, can effectively modulate neuronal excitability.
Understanding the intricate structure of membrane proteins is pivotal, especially in neurological disorders like epilepsy, autism, and schizophrenia, which are involved in dysfunction of KCC2. Our study explored the structure, function, and significance of KCC2 in neurological health and disorders, highlighting its importance in both normal neuronal activity and various pathologies.
By using our proprietary innovative detergent, we successfully solubilized and purified native, unaggregated KCC2, shedding light on its structure. Electron microscopy revealed that KCC2 exists in both monomeric and dimeric forms, with monomers displaying distinct "head" and "core" domains connected by a flexible linker. The dimers exhibited an asymmetrical S-shaped architecture, with disulfide bridges playing a crucial role in dimerization. Mass spectrometry identified specific post-translational modifications, including glycosylations and phosphorylations, which align with KCC2's functional profile. Notably, specific binding with an inhibitor (VU0463271) was confirmed through surface plasmon resonance (SPR), allowing to verify its functional integrity. These structural insights were complemented by functional studies demonstrating that adding a tag to the C-terminus impaired KCC2 function. This highlights the critical nature of this region. The glycosylations and phosphorylations, aligning with KCC2's functional profile are also revealed by mass spectrometry (MS).
Overall, these results have provided a comprehensive view of KCC2's structure and function relationship, offering a solid foundation for future research into neurological disorders and potential therapeutic interventions.
Unlock the potential of your research with Eurofins CALIXAR’s innovative solutions for membrane proteins, including KCC2 and various ion channels. Our high-purity, biologically active proteins are ideal for applications like high-throughput screening, antibody development, and structural biology studies. Partner with us to accelerate your drug development program and explore new therapeutics for neurological disorders and other conditions linked to ion channel dysfunction. Elevate your research today!
Explore our case studies and discover how Eurofins CALIXAR can contribute to your next scientific breakthrough. For custom inquiries or collaborations, please contact us.
References :
Desuzinges Mandon E. et al. 2017
