Post-translational histone modifications associated with the development of metabolic dysfunction-associated fatty liver disease. Part 1. General provisions

GASTROENTEROLOGY, Journal Year: 2024, Volume and Issue: 58(3), P. 210 - 221

Published: Sept. 13, 2024

Based on the analysis of literary sources PubMed, MedLine, The Cochrane Library, EMBASE database, authors article give general provisions regarding post-translational modifications histones (small proteins with a molecular weight 10–15 kDa, which make up largest part nuclear proteins), are associated development metabolic dysfunction-associated fatty liver disease. emphasize that histone regulate activity gene expression, and each these types differently changes structure chromatin and, as result, expression. Currently, more than 20 protein have been identified (acetylation, biotinylation, butyrylation, 2-hydroxybutyrylation, ADP-ribosylation, N-formylation, hydroxylation, glycosylation, glutarylation, dopaminylation, proline isomerization aspartic acid carbonylation, crotonylation, lactylation, malonylation, methylation, propionylation, succinylation, SUMOylation, ubiquitination, phosphorylation, citrullination). Epigenetic epitranscriptomic induced by lifestyle, especially nature diet physical activity, influence exogenous endogenous factors. Prolonged epigenetic determine expression target genes can be accompanied disorders progression Histone modification is carried out site-specific enzymes: writers, identify marker, erasers, “erase” marker. Post-translational change local physicochemical environment based this, directly affect nucleosome chromatin. Also, N- C-terminal tails act “docking sites” recruit specific readers. Readers both in intranucleosomal space, modifying adjacent sites or recruiting transcription factors, activators repressors, internucleosomal space. also describe pathophysiological significance disease, diagnostic value biomarkers, potential pharmacological management to achieve inhibition pathological process.

Language: Английский

Sensitive Detection of Histones and γ-H2AX by Immunoblotting: Problems and Solutions

Chemical Research in Toxicology, Journal Year: 2024, Volume and Issue: 37(9), P. 1588 - 1597

Published: Sept. 5, 2024

Histones and their posttranslational modifications (PTMs) are critical regulators of gene expression. Differentiation, environmental stressors, xenobiotics, major human diseases cause significant changes in histone variants PTMs. Western blotting is the mainstay methodology for detection histones PTMs majority studies. Surprisingly, despite high abundance cells, immunoblotting typically involves loading large protein amounts that normally used sparse cellular proteins. We systematically examined technical factors Western-blotting-based with >30 antibodies. found under multiple transfer conditions, many epitopes on polyvinylidene fluoride (PVDF) membranes had a very low antibody accessibility, which was dramatically increased by addition simple denaturation step. Denaturation membrane-bound proteins also enhanced specificity some In comparison to standard PVDF membranes, sensitivity nitrocellulose much higher, further inclusion same Optimized protocols >100-times genotoxic marker γ-H2AX two monoclonal The impact use varied different histones, but each histone, it generally similar antibodies targeting N-terminal C-terminal regions. summary, strongly improves Westerns, resulting more accurate measurements permitting analyses small biological samples.

Language: Английский