Thiol post-translational modifications modulate allosteric regulation of the OpcA-G6PDH complex through conformational gate control

TL;DR

This study uncovers how thiol post-translational modifications of OpcA regulate G6PDH activity through allosteric conformational changes, enabling cyanobacteria to rapidly adapt their metabolism to environmental changes.

Contribution

It reveals the molecular mechanism by which thiol PTMs of OpcA modulate G6PDH allosterically, combining computational modeling and redox proteomics.

Findings

Thiol PTMs near the OpcA-G6PDH interface are crucial for allosteric regulation.

PTMs enhance G6PDH activity during oxidation of OpcA.

The study demonstrates PTM-driven rapid metabolic switching in cyanobacteria.

Abstract

Cyanobacteria require ultra-fast metabolic switching to maintain reducing power balance during environmental fluctuations. Glucose-6-phosphate dehydrogenase (G6PDH), catalyzing the rate-limiting step of the oxidative pentose phosphate pathway (OPPP), provides essential NADPH and metabolic intermediates for biosynthetic processes and redox homeostasis. In cyanobacteria, the unique redox-sensitive protein OpcA acts as a metabolic switch for G6PDH, enabling rapid adjustment of reducing power generation from glycogen catabolism and resulting in precise regulation of carbon flux between anabolic and catabolic pathways. While the redox-sensitive cysteine structures of OpcA are known to regulate G6PDH, the detailed mechanisms of how redox post-translational modifications (PTMs) influence OpcA's allosteric effects on G6PDH structures and function remain elusive. To investigate this mechanism, we utilized computational modeling combined with experimental redox proteomics using Synechococcus elongatus PCC 7942 as a model system. Redox proteomics captured modified cysteine residues under light/dark or circadian shifts. Computational simulation revealed that thiol PTMs near the OpcA-G6PDH interface are crucial to allosteric regulation of regions affecting the G6PDH activity, including a potential gate region for substrate ingress and product egress, as well as critical hydrogen bond networks within the active site. These PTMs promote rapid metabolic switching by enhancing G6PDH catalytic activity when OpcA is oxidized. This study provides evidence for novel molecular mechanisms that elucidate the importance of thiol PTMs of OpcA in modulating G6PDH structure and function in an allosteric manner, demonstrating how PTM-level regulation provides a critical control mechanism that enables cyanobacteria to rapidly adapt to environmental fluctuations through precise metabolic fine-tuning.