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. Author manuscript; available in PMC: 2022 Jun 20.
Published in final edited form as: Annu Rev Biochem. 2021 Jun 20;90:287–320. doi: 10.1146/annurev-biochem-080120-021109

Figure 7.

Figure 7

Protein semisynthesis and characterization of ubiquitylated H2B. (a) Synthesis of H2BK120ub peptides using an aminothiol ligation auxiliary. (b) Semisynthesis of the full-length, native H2BK120ub protein via two-step traceless ligation route. (c) Streamlined synthesis of full-length H2BK120ub with a G76A ubiquitin modification. (d) Structure of the disulfide-linked ubiquitin analog, H2B-120ubSS. (e) Structure of Dot1L engaging a nucleosome with the H2BK120ub modification. Residues mediating the Dot1L-ubiquitin interaction are shown. PDB, 6nn6. (f) Quantification of chromatin array compaction with homo-FRET, measured by changes in steady-state anisotropy (SSA). (g) Probing ubiquitin interactions in compacted chromatin arrays with H/D exchange and NMR spectroscopy. PGs, protecting groups. MesNa, sodium 2-mercaptoethanesulfonate. TCEP, tris(2-carboxyethyl)phosphine.