Metabolite interactions in the bacterial Calvin cycle and implications for flux regulation

ABSTRACT: Metabolite-level regulation of enzyme activity is important for microbes to cope with environmental shifts. Knowledge of such regulations can also guide strain engineering to improve industrial phenotypes. Recently developed chemoproteomics workflows allow for genome-wide detection of metabolite-protein interactions that may regulate pathway activity. We applied limited proteolysis small molecule mapping (LiP-SMap) to identify and compare metabolite-protein interactions in the proteomes of two cyanobacteria and two lithoautotrophic bacteria that fix CO2 using the Calvin cycle. Clustering analysis of the hundreds of detected interactions showed that some metabolites interacted in a species-specific manner, such as interactions of glucose-6-phosphate in Cupriavidus necator and of glyoxylate in Synechocystis sp.sp PCC 6803. These are interpreted in light of the different central carbon conversion pathways present. Metabolites interacting with the Calvin cycle enzymes fructose-1,6/sedoheptulose-1,7-bisphosphatase (F/SBPase) and transketolase were tested for effects on catalytic activity in vitro. The Calvin cycle intermediate glyceraldehyde-3-phosphate activated both Synechocystis and Cupriavidus F/SBPase, which suggests a feed-forward activation of the cycle in both photoautotrophs and chemolithoautotrophs. In contrast to the stimulating effect in reduced conditions, glyceraldehyde-3-phosphate inactivated the Synechocystis F/SBPase in oxidized conditions by accelerating protein aggregation. Thus, metabolite-level regulation of the Calvin cycle is more prevalent than previously appreciated and may act in addition to redox regulation.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Synechococcus Elongatus (strain Pcc 7942) (anacystis Nidulans R2) Synechocystis Sp. (strain Pcc 6803 / Kazusa) Cupriavidus Necator (strain Atcc 17699 / H16 / Dsm 428 / Stanier 337) (ralstonia Eutropha) Bacteria Hydrogenophaga Pseudoflava

SUBMITTER: Emil Sporre  

LAB HEAD: Elton Paul Hudson

PROVIDER: PXD044412 | Pride | 2023-08-23

REPOSITORIES: Pride