XC
Xiongwen Cao
  • Post-Doc, Yale university New Haven
Research fields
  • Biochemistry, Cancer Biology, Cell Biology, Molecular Biology, Neuroscience
BONCAT-based Profiling of Nascent Small and Alternative Open Reading Frame-encoded Proteins
Authors:  Xiongwen Cao, Yanran Chen, Alexandra Khitun and Sarah A. Slavoff, date: 01/05/2023, view: 924, Q&A: 0

RIBO-seq and proteogenomics have revealed that mammalian genomes harbor thousands of unannotated small and alternative open reading frames (smORFs, <100 amino acids, and alt-ORFs, >100 amino acids, respectively). Several dozen mammalian smORF-encoded proteins (SEPs) and alt-ORF-encoded proteins (alt-proteins) have been shown to play important biological roles, while the overwhelming majority of smORFs and alt-ORFs remain uncharacterized, particularly at the molecular level. Functional proteomics has the potential to reveal key properties of unannotated SEPs and alt-proteins in high throughput, and an approach to identify SEPs and alt-proteins undergoing regulated synthesis should be of broad utility. Here, we introduce a chemoproteomic pipeline based on bio-orthogonal non-canonical amino acid tagging (BONCAT) (Dieterich et al., 2006) to profile nascent SEPs and alt-proteins in human cells. This approach is able to identify cellular stress-induced and cell-cycle regulated SEPs and alt-proteins in cells.


Graphical abstract



Schematic overview of BONCAT-based chemoproteomic profiling of nascent, unannotated small and alternative open reading frame-encoded proteins (SEPs and alt-proteins)

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