Ocean acidification remodels the mantle phosphoproteome and weakens shell mechanical resilience in Mytilus edulis.

Ocean acidification (OA), driven by rising atmospheric CO2, threatens the structural integrity and ecological performance of calcifying marine organisms, yet the mechanistic links between environmental acidification and compromised shell mechanical resilience remain insufficiently understood. Here, we integrate microstructural imaging, mechanical testing, and quantitative phosphoproteomics to characterize the structural, mechanical, and phosphorylation responses of Mytilus edulis to OA and examine their relationships. Adult mussels were exposed to near-future pH conditions (8.1, 7.9, 7.7) for up to 40 days. OA produced progressive disorganization of nacreous tablets and prismatic columns without altering aragonite-calcite polymorphs, indicating that acidification drives structural degradation rather than mineralogical shifts. Correspondingly, shell compressive strength declined in a pH- and time-dependent manner, with an approximately 60% reduction at pH 7.7 after 40 days. Phosphoproteomic profiling identified 3,720 phosphoproteins (10,780 sites), revealing extensive acidification-induced remodeling of phosphorylation networks. Proteins corresponding to differentially phosphorylated peptides were predominantly nuclear and were enriched in kinase-, PH-, PDZ-, and RNA-recognition-domain-containing proteins. Functional analyses implicated MAPK signaling, vesicular trafficking, ion transport, glycolysis/gluconeogenesis, autophagy, and calcium-associated processes. Phosphoproteomic remodeling was associated with the deterioration of shell microstructure and compressive strength, supporting an association between mantle phosphorylation responses and compromised biomineralization under acidification. The enriched pathways and protein domains were functionally related to calcium-dependent signaling, vesicular trafficking, cytoskeletal organization, and cellular metabolism, providing testable candidates for future mechanistic validation. Together, these findings support a working model in which ocean acidification is accompanied by coordinated phosphorylation remodeling and progressive loss of shell mechanical resilience. We further propose a proof-of-concept dual-indicator framework that integrates candidate molecular sentinels with organismal metrics, providing a conceptual basis for future assessment of aquaculture resilience and ecosystem vulnerability under progressive ocean acidification.