In-cell processing enables rapid and in-depth proteome analysis of low-input <i>Caenorhabditis elegans</i>.

ABSTRACT: Caenorhabditis elegans is a widely used genetic model organism, however, the worm cuticle complicates extraction of intracellular proteins, a prerequisite for typical bottom-up proteomics. Conventional physical disruption procedures are not only time-consuming, but can also cause significant sample loss, making it difficult to perform proteomics with low-input samples. Here, for the first time, we present an on-filter in-cell (OFIC) processing approach, which can digest C. elegans proteins directly in the cells of the organism after methanol fixation. With OFIC processing and single-shot LCMS analysis, we identified over 9,400 proteins from a sample of only 200 worms, the largest C. elegans proteome reported to date that did not require fractionation or enrichment. We

SUBMITTER: Elsayyid M 

PROVIDER: S-EPMC11429863 | biostudies-literature | 2024 Sep

REPOSITORIES: biostudies-literature