Fine‐Tuning Protein Self‐Organization by Orthogonal Chemo‐Optogenetic Tools

다국어 초록 (Multilingual Abstract)

A universal gain‐of‐function approach for the spatiotemporal control of protein activity is highly desirable when reconstituting biological modules in vitro. Here we used orthogonal translation with a photocaged amino acid to map and elucidate molecular mechanisms in the self‐organization of the prokaryotic filamentous cell‐division protein (FtsZ) that is highly relevant for the assembly of the division ring in bacteria. We masked a tyrosine residue of FtsZ by site‐specific incorporation of a photocaged tyrosine analogue. While the mutant still shows self‐assembly into filaments, dynamic self‐organization into ring patterns can no longer be observed. UV‐mediated uncaging revealed that tyrosine 222 is essential for the regulation of the protein's GTPase activity, self‐organization, and treadmilling dynamics. Thus, the light‐mediated assembly of functional protein modules appears to be a promising minimal‐regulation strategy for building up molecular complexity towards a minimal cell.
We developed orthogonal chemo‐optogenetic tools as a minimal regulation strategy to fine‐tune in‐vitro protein self‐organization with light. Our chemo‐optogenetic tools are designed by orthogonal translation with an expanded genetic code. Such introduced bio‐orthogonal chemical groups operate at the level of single specific amino acids, but perform tight control of enzyme activity and, thereby, the ability of protein self‐organization.