Proteome profile of peripheral myelin in healthy mice and in a neuropathy model

Using the sciatic nerve as a model, we biochemically purified peripheral nervous system (PNS) myelin and systematically assessed its proteome by dedicated, partly ion mobility-enhanced data-independent acquisition (DIA) workflows with alternating low and elevated energy (MSE) modes. Specifically, we used MSE for correct quantification of highest-abundance proteins, UDMSE for deep quantitative proteome coverage, and dynamic range enhancement (DRE)-UDMSE for comparative analysis, the latter mode providing a good compromise between dynamic range, identification rate and instrument run time for routine differential myelin proteome profiling. We found that integration of the proteome data with the total nerve transcriptome is suited to determine comprehensive molecular profiles in healthy nerves and in myelin-related disorders. This work provides the most comprehensive proteome resource thus far to approach development, function and pathology of peripheral myelin, and a straightforward, accurate and sensitive workflow to address myelin diversity in health and disease.