Fine-Tuning Protein Self-Organization by Orthogonal Chemo-Optogenetic Tools.

Huan Sun,Nediljko Budisa,Diego A Ramirez‐Diaz,Petra Schwille,Haiyang Jia

doi:10.1002/anie.202008691

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.

Highlights

A universal gain-of-function approach for the spatiotemporal control of protein activity is highly desirable when reconstituting biological modules in vitro
The reconstituted biological system we focused on is the most well-known prokaryotic division protein, filamenting temperature-sensitive mutant Z, abbreviated FtsZ, which is widely conserved in bacteria and
We find the new fidelity of ONBY incorporation into FtsZ was verified by ESI-MS

Summary

Introduction

A universal gain-of-function approach for the spatiotemporal control of protein activity is highly desirable when reconstituting biological modules in vitro. As a consequence, photocaged FtsZ self-organizes into dynamic ring formation in membrane. FtsZ(Y222F)-YFP-mts demonstrates a similar GTPase activity compared to WT, indicating that removing a hydroxyl group will not reduce the GTP hydrolysis capability.

Results

Conclusion