in-vitro-reconstitution-self-organizing-protein-patterns-on-supported

<em>In Vitro</em> Reconstitution of Self-Organizing Protein Patterns on Supported Lipid Bilayers

Many aspects of the fundamental spatiotemporal organization of cells are governed by reaction-diffusion type systems. In vitro reconstitution of such ...

Max-Planck-Institut für Biochemie

Max Planck Institute of Biochemistry

The MinDE protein system from E. coli has recently been identified as a minimal biological oscillator, based on two proteins only: The ATPase MinD and the ATPase activating protein MinE. In E. coli, the system works as the molecular ruler to place the divisome at midcell for cell division. Despite its compositional simplicity, the molecular mechanism leading to protein patterns and oscillations is still insufficiently understood. Here we used high-speed atomic force microscopy to analyze the mechanism of MinDE membrane association/dissociation dynamics on isolated membrane patches, down to the level of individual point oscillators. This nanoscale analysis shows that MinD association to and dissociation from the membrane are both highly cooperative but mechanistically different processes. We propose that they represent the two directions of a single allosteric switch leading to MinD filament formation and depolymerization. Association/dissociation are separated by rather long apparently silent periods. The membrane-associated period is characterized by MinD filament multivalent binding, avidity, while the dissociated period is defined by seeding of individual MinD. Analyzing association/dissociation kinetics with varying MinD and MinE concentrations and dependent on membrane patch size allowed us to disentangle the essential dynamic variables of the MinDE oscillation cycle.

Beatrice Ramm

Department of Cellular and Molecular Biophysics

Max-Planck-Institut für Biochemie

In Vitro Reconstitution of Self-Organizing Protein Patterns on Supported Lipid Bilayers

Beatrice Ramm

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Cellular and Molecular Biophysics

Max-Planck-Institut für Biochemie

In Vitro Reconstitution of Self-Organizing Protein Patterns on Supported Lipid Bilayers

Department of Cellular and Molecular Biophysics