Lipid vesicle binding and modulation of membrane binding proteins and compounds

The lipid bilayer is an essential component of cells that separates the internal components to the external environment and also defines the internal components. Various proteins are utilized by the membrane for various cell processes including signaling, communication and responding to external signals. The interaction between cell membranes and proteins may have biological consequences [Lemmon MA, 2008 ], so studying these interactions allows a deeper understanding of biological processes and may reveal the pathogenesis of diseases related to these proteins.

Class 1 Myosins are a group of actin based motor proteins associated with various roles related to membrane dynamics and trafficking. Their tail domain contains membrane binding regions such as the TH1 domain and a pleckstrin homology (PH) domain within the TH1.

Alpha Synuclein is a predominantly presynaptic neuronal protein linked to several neurodegenerative disorders such as Parkinson's disease. It has been shown to interact with membranes and to associate with synaptic vesicles. In its physiological state, it has been shown to be acetylated, however the effect of this modification is still being investigated.

Bacterial resistance to antimicrobials is becoming a significant challenge in effective treatment and prevention of bacterial infections worldwide, and the need for new antimicrobials is rising. For antimicrobials to be successful, they must initially penetrate and interact with the bacterial membranes in order to carry out their function.

This project focused on the membrane binding effects of Myosin 1 and alpha Synuclein in order to determine their function and whether post-translational modifications impact these interactions. We also looked at antibiotic compounds to determine their membrane interactions using lipid vesicles as a model for the cell membrane and analyzing the interaction using stopped flow, DLS (dynamic light scattering) data and microscopy images. We successfully cloned and expressed both forms of the TH1 domain and were able to analyze the interaction between alpha Synuclein and vesicles. Our results suggest that the lipid composition of the vesicles affects their interaction with alpha Synuclein. Furthermore, acetylation of alpha Synuclein alters these interactions.