Journal: Nature

Article Title: Structural mechanism of angiogenin activation by the ribosome.

doi: 10.1038/s41586-024-07508-8

Figure Lengend Snippet: Fig. 1 | Angiogenin inhibits translation by binding to 80S ribosomes. a, Angiogenin (Ang) inhibits translation in RRL in a concentration-dependent manner. Inhibition is relieved by RNasin (RNase inhibitor). The apparent translation rate is shown (relative light units (RLU) s−1). Data are mean ± s.e.m. n = 2 or 3 independent experiments, as shown. b, Incubation of RRL with angiogenin for 15 min leads to the accumulation of tRNA fragments; fragment accumulation is inhibited by RNasin. n = 2 independent experiments. nt, nucleotides. c, Angiogenin co-pellets with the ribosome fraction from RRL, except in the presence of RNasin. n = 2 independent experiments. d, Cryo-EM analysis of RRL with 1 µM angiogenin reveals that over 30% of 80S ribosomes have a novel density in the A site. e, Rigid-body fit of human angiogenin (PDB: 5EOP)61 into the 3.4 Å cryo-EM map of angiogenin-bound ribosomes in RRL. f, A 3.0 Å cryo-EM map of in vitro assembled 80S–angiogenin complex. The 60S subunit is coloured cyan; 40S subunit is coloured yellow; P-site tRNA is coloured

Article Snippet: For anti-RNase A blotting, procedures were amended to block with 1× PBST with 0.1 mg ml−1 BSA and detected with 1:1,000 rabbit anti-bovine pancreatic RNase A polyclonal antibody-peroxidase conjugated (200-4388S, Rockland Immunochemicals).

Techniques: Binding Assay, Concentration Assay, Inhibition, Incubation, Cryo-EM Sample Prep, In Vitro