Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a debilitating disease with unknown causes. It is recognized as a multi-systemic disorder involving immune dysfunction, mitochondrial impairment, neuroinflammation, autonomic dysregulation, and metabolic abnormalities. Protein disorder is characterized by increased accessibility, structural flexibility, and dynamic instability, which can lead to widespread cellular dysfunction and contribute to various diseases, including neurodegenerative diseases. While the underlying mechanisms of ME/CFS remain elusive, studies have suggested the involvement of disruptions in protein folding, clearance, and degradation, linking them closely to protein disorder and further inspiring our exploration of their relationship.
To gain new insights into the mechanisms of ME/CFS, in this study, we aim to utilize an innovative proteomic workflow using a unique chemical probe, TME, to profile protein disorder. TME specifically labels proteins with free cysteines located in surface-exposed and conformationally flexible environments—a hallmark of protein unfolding and disordered regions—while its alkyne group enables capture and enrichment of disordered proteins. We first established a new workflow that allows site-specific modifications of proteins to be identified using TME. This workflow will then be applied to identify and quantify disordered regions in the proteome of fibroblast cells from individuals with ME/CFS and healthy controls. Further enrichment analysis will be performed with the goal to provide a comprehensive overview of disordered proteins in ME/CFS, advancing our understanding of ME/CFS pathophysiology and improving its diagnosis and treatment.