ArticleMethods in molecular biology (Clifton, N.J.)2026
Proteomic Detection of Small and Alternative Open Reading Frame-Encoded Microproteins and Alternative Proteins.
Article in Methods in molecular biology (Clifton, N.J.), 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
RIBO-seq and proteomics have revealed that mammalian genomes harbor thousands of unannotated small and alternative open reading frames (sm/alt-ORFs <150 codons) that are translated into microproteins and alternative proteins (micro/alt-proteins). Several dozens of micro/alt-proteins have been characterized at the molecular level and demonstrated to play important biological roles. However, the overwhelming majority of micro/alt-proteins remain undefined, possibly because they are excluded from protein databases and cannot be detected by a classical proteomic workflow. Here, we introduce a proteomic pipeline to detect unannotated micro/alt-proteins in cultured cells or tissues from total protein extraction, gel-based size selection, digestion, and liquid chromatography-mass spectrometry proteomics (LC-MS/MS), to data analysis and manual validation. This approach is able to identify unannotated micro/alt-proteins from cultured cells and (patho)physiological tissues.
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