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bioRxiv · 10.1101/2024.07.05.602292

Charge distribution and helical content tune the binding of septins amphipathic helix domain to lipid membranes

Edelmaier, C. J.·Klawa, S. J.·Mofidi, S. M.·Wang, Q.·Bhonge, S.·Vogt, E. J. D.·Curtis, B. N.·Shi, W.·Hanson, S. M.·Klotsa, D.·Forest, M. G.·Gladfelter, A. S.·Freeman, R.·Nazockdast, E.
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Abstract

Septins are a class of cytoskeletal proteins that preferentially bind to domains of micron-scale curvature on the cell membrane. Studies have shown that amphipathic helix (AH) domains in septin oligomers are essential for septin curvature sensing. Yet, the underlying mechanochemical interactions that modulate this curvature sensing remain ambiguous. Here we use all-atom molecular dynamics alongside a metadynamics enhanced sampling approach to bridge the gap between time and length scales required to optimize and validate experimental design of amphipathic helices. Simulations revealed that the local charge on the termini of an 18-amino-acid AH peptide impacts its helical content and positioning within lipid membranes. These computational observations are confirmed with experiments measuring the binding of synthetic AH constructs with variable helical content and charged termini to lipid vesicles. Taken together, these results identify the helical content of amphipathic helices as a regulator of septin binding affinity to lipid membranes. Additionally, we examined an extended AH sequence including 8 amino acids upstream and downstream of the minimal 18-amino-acid-long AH domain to more closely mimic the native protein in simulations and experiments. Simulations and experiments show that the extended peptide sequence adopts a strong alpha-helical conformation when free in solution, giving rise to a higher affinity to lipid membranes than that of the shorter AH sequence. Together, these results provide insight into how the native septin proteins interact with membranes, and establish general design principles that can guide the interaction of future synthetic materials with lipid membranes in a programmable manner. STATEMENT OF SIGNIFICANCEUnderstanding how cells sense and react to their shape is necessary for numerous biological processes. Here we explore the interactions between amphipathic helices, a curvature sensing protein motif, and lipid membranes. Using molecular dynamics simulations, enhanced simulation sampling techniques, and experiments, we find that increasing the helical content of the amphipathic helix or adding charged capping sequences yields higher membrane binding affinity. Understanding these parameters for membrane-binding could enable us to interface and regulate native protein functions, as well as guide the design of synthetic curvature-sensing materials that can interact with and deform lipid membranes.

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Edelmaier, C. J., Klawa, S. J., Mofidi, S. M., Wang, Q., Bhonge, S., Vogt, E. J. D., Curtis, B. N., Shi, W., Hanson, S. M., Klotsa, D., Forest, M. G., Gladfelter, A. S., Freeman, R., Nazockdast, E.. 2024-07-10. Charge distribution and helical content tune the binding of septins amphipathic helix domain to lipid membranes. https://doi.org/10.1101/2024.07.05.602292

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