METHODOLOGICAL TOOLBOX FOR DETECTION AND RESEARCH OF MICROPEPTIDES: FROM GENOME TO FUNCTION

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Resumo

Micropeptides encoded by small open reading frames (sORFs) are an emerging class of functional molecules that regulate key cellular processes. Their study is hampered by several methodological challenges, including their small size, low abundance, and the difficulty in generating specific antibodies. This review systematizes current approaches for the identification and functional characterization of micropeptides. We describe the main strategies for their discovery: bioinformatic algorithms, global translation analysis via ribosome profiling, direct detection by mass spectrometry-based proteomics, and phenotypic screenings. Additionally, methods for functional validation and the elucidation of their molecular mechanisms of action are reviewed, including genetic knockouts and affinity tagging for visualization and the study of protein-protein interactions. The review discusses the key challenges and future prospects of the field, emphasizing the importance of an integrated multi-omics approach for the comprehensive mapping of the micropeptidome.

Sobre autores

A. Lavrov

Lomonosov Moscow State University; Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University

119991 Moscow, Russia; 119992 Moscow, Russia

N. Shepelev

Lomonosov Moscow State University; Shennyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences

119991 Moscow, Russia; 117997 Moscow, Russia

O. Dontsova

Lomonosov Moscow State University; Shennyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences

119991 Moscow, Russia; 117997 Moscow, Russia

M. Rubtsova

Lomonosov Moscow State University; Shennyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences

Email: mprubtsova@gmail.com
Correspondence address 119991 Moscow, Russia; 117997 Moscow, Russia

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