Lipid–protein interactions are central to maintaining the structural and functional balance of biological membranes, influencing a wide array of cellular processes. These interactions, however, become pathological in neurodegenerative diseases (NDDs), such as Alzheimer’s, Parkinson’s, and Huntington’s diseases. In these disorders, the misfolding and aggregation of proteins like amyloid-beta (Aβ), alphasynuclein (α-syn), and mutant huntingtin (mHTT) disrupt the lipid bilayer, compromising membrane integrity, fluidity, and signaling. In this review we explore the critical role of lipid–protein interactions in NDDs, emphasizing how protein misfolding leads to toxic aggregates that embed into membranes, triggering neurotoxic events. Advanced spectroscopic techniques have been instrumental in studying these molecular interactions. Photon-based methods, including Förster resonance energy transfer (FRET), circular dichroism (CD), and Raman spectroscopy, provide real-time insights into protein aggregation and lipid membrane dynamics. Neutron-based techniques, such as neutron reflectometry and small-angle neutron scattering (SANS), further enhance the resolution of lipid–protein interactions, particularly in the context of neurodegenerative aggregation.
Moreover, the review highlights the significance of lipid microdomains, particularly cholesterol-rich lipid rafts, which act as platforms for protein aggregation, influencing disease progression. Therapeutic strategies aimed at targeting these lipid–protein interfaces are also discussed, with a focus on how spectroscopic insights have driven the development of drugs that stabilize membrane integrity or prevent toxic aggregation. Finally, the integration of spectroscopy with computational models, such as molecular dynamics (MD) simulations, is proposed as a promising approach to further unravel the complex dynamics of lipid–protein interactions, providing a more complete picture of disease mechanisms.
This study provides a comparative analysis of various components of mesenchymal stem cells (MSC) conditioned media (CM) obtained using serum-containing and serum-free culture methods, revealing significant differences in their composition and potential clinical applicability. Serum-containing CM exhibits significantly higher levels of total protein, non-vesicular RNA, exosomes, and nanoparticles compared to serum-free CM, reflecting the contribution of both the MSC secretome and residual fetal bovine serum components. Ultrafiltration-based fractionation (0.2 µm–50 kDa) allows the isolation of fraction enriched in exosomes and proteins, preserving the functionally significant components of the MSC secretome. This strategy effectively captures small vesicles and mid-sized proteins while excluding larger or smaller biomolecules, enhancing utility for targeted analyses. The presented data underscore the need for context-driven CM selection and provide information for choosing the optimal strategy for obtaining the MSC secretome balancing yield, purity, and regulatory demands in MSC research and therapy.
