Structural and systems characterization of phosphorylation on metabolic enzymes identifies sex-specific metabolic reprogramming in obesity

ABSTRACT: The coordination of cellular signaling networks with metabolic response is key for balanced energy production and homeostasis. This coordination is achieved through spatiotemporal control of metabolism via compartmentalization and redundancies coupled to rapid signaling. Such dynamics must therefore require fast regulatory networks such as those directed by phosphorylation of serine (S) (~90%), threonine (T) (~9%), and tyrosine (Y) (~0.1-1%) residues on metabolic enzymes. In order to determine the structure-function relationship of phosphorylation sites on metabolic enzymes, we leveraged the published phosphoproteome and structural data for metabolic enzymes to stratify phosphorylation sites in the context of functional domains. There was significant enrichment of pY in proximity to functional and dimerization domains.In order to gain network level insight into the pY directed regulation of metabolic enzymes and the resulting dynamics of metabolic reprogramming, we employed proteomics, metabolomics, structural analysis, and computational modeling to characterize the functional impact of pY on enzymes in the context of obesity. We validated the intrinsic role of select phosphosites on enzyme function via enzyme kinetics assays and isotope labeled metabolic tracing. Overall, our multidisciplinary approach bridges the structure-function knowledge gap allowing us to identify the convergence zone where cellular signaling ‘tunes’ metabolism.

INSTRUMENT(S): Q Exactive Plus, Orbitrap Exploris 480

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Liver

DISEASE(S): Obesity

SUBMITTER: Tigist Tamir  

LAB HEAD: Forest White

PROVIDER: PXD054497 | Pride | 2025-05-17

REPOSITORIES: Pride