This detailed volume brings together cutting-edge methods from proteomics, transcriptomics, bioinformatics, and functional genomics in order to capture the dynamic and multidisciplinary nature of the microprotein field. Divided into four sections, the book explores the detection and identification of microproteins; transcriptome-based discovery; computational, structural, and evolutionary insights; and functional and biomedical relevance. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.

Authoritative and practical, Microproteins: Methods and Protocols serves not only as a technical guide but also as an inspiration for further exploration into these once-hidden elements of the proteome.

Inhalt

Microproteins: Uncovering Hidden Layers of the Proteome.- Proteomic Detection of Small and Alternative Open Reading Frame-Encoded Microproteins and Alternative Proteins.- Identification of Canonical and Alternative Proteins by Mass Spectrometry.- Identification and Characterization of Microproteins/MicroORFs.- Proteogenomic Identification of Annotated and Novel smORFs via Microprotein Enrichment.- Identification of Peptides Encoded by Small Open Reading Frames in Microalgae by Mass Spectrometry.- Identifying and Visualizing Novel Small Open Reading Frames (sORFs) via Ribo-Seq, Transcriptome Assembly, and ggRibo.- Translation Signature Scores: Data-Driven Approach to Assess Evidence for Active Translation.- Identification of Potentially Coding Small ORFs in Plant Transcriptomes.- Structure and Disorder Predictions of Microproteins: Usage, Applications, and Pitfalls.- Computational Methods for Tracing the Evolutionary History of Human Microproteins Encoded by Intronless Genes.- Bioinformatic Tools for Non-Bioinformaticians: A Roadmap to Study Unknown Microprotein Sequences.- Functional Testing of Microproteins in a Vertebrate Model of Development.- Functional Validation of Transcriptionally Active Microproteins/sORFs In Planta.- Elucidating the Localization and Interactome of Mitochondrial Microproteins.- Illuminating Small Proteins through HiBiT Blotting.- Gain-of-Functional Genomic Screening for Microproteins Essential for Tumorigenesis.- Proteomic Identification of Microproteins Translated from Long Non-Coding RNA.- Urinary Micropeptides: Integrative In Silico Profiling and LC-MS/MS Analysis for Prostate Cancer Biomarker Discovery.- Using Untargeted Mass Spectrometry-Based Proteomics to Identify Putative Microprotein Biomarkers from Patient Tampons.