PROJECT SUMMARY Although the health benefits of physical activity in the prevention and mitigation of many chronic diseases have been documented for decades, the molecular mechanisms mediating these health benefits are still poorly understood. To understand the adaptive response of physical activity at the molecular level, it is essential to comprehensively characterize the dynamic changes in protein expression and post-translational modifications (PTMs) in. The overall objective of the PNNL Proteomics Chemical Analysis Center is to support the consortium by providing comprehensive discovery of proteins and associated PTMs in skeletal muscle and subcutaneous adipose tissues collected by Clinical Centers of MoTrPAC. To aid in the development of a ‘molecular map’ of transducers that provides the critical insights and molecular impacts of physical activity in humans, our integrative proteomics analysis plan focuses on two major tissues known to be important in exercise signaling especially with training adaptations. This work will intracellular signaling as mediated by protein PTMs, including protein phosphorylation, lysine acetylation, and pilot studies of reversible redox modifications. The High level summary of the Specific Aims of this project are: 1) Analyze and publish expansive multi-omics datasets in a collaborative manner. 2) Collaborate with CC investigators and the rest of MoTrPAC to implement temporal multi-omics profiling of muscle and adipose tissue following an acute exercise bout in highly active adults. 3) Perform state-of-the-art proteomics profiling based on the human tissue analysis plan. 4) Coordinate with other MoTrPAC activities including efforts under the CCC, inclusive of the 1) Administrative Coordinating Core (ACC), 2) Data Management, Analysis, and Quality Control Core (DMAQC), 3) Exercise Intervention Core (EIC), and 4) Biological Sample Repository (BSR). 5) Participate on Committees, Subcommittees, and Working Groups to facilitate communications, coordinate activities across all components of MoTrPAC, conduct continuous quality assurance of data, develop analytical strategies, and prepare presentations and manuscripts to disseminate results.