Summary information and primary citation
- PDB-id
- 7kwg; SNAP-derived features in text and JSON formats;
DNAproDB
- Class
- ribosome
- Method
- cryo-EM (3.75 Å)
- Summary
- Staphylococcus aureus 30s ribosomal subunit in presence of spermidine
- Reference
- Belinite M, Khusainov I, Soufari H, Marzi S, Romby P, Yusupov M, Hashem Y (2021): "Stabilization of Ribosomal RNA of the Small Subunit by Spermidine in Staphylococcus aureus ." Front Mol Biosci, 8, 738752. doi: 10.3389/fmolb.2021.738752.
- Abstract
- Cryo-electron microscopy is now used as a method of choice in structural biology for studying protein synthesis, a process mediated by the ribosome machinery. In order to achieve high-resolution structures using this approach, one needs to obtain homogeneous and stable samples, which requires optimization of ribosome purification in a species-dependent manner. This is especially critical for the bacterial small ribosomal subunit that tends to be unstable in the absence of ligands. Here, we report a protocol for purification of stable 30 S from the Gram-positive bacterium Staphylococcus aureus and its cryo-EM structures: in presence of spermidine at a resolution ranging between 3.4 and 3.6 Å and in its absence at 5.3 Å. Using biochemical characterization and cryo-EM, we demonstrate the importance of spermidine for stabilization of the 30 S via preserving favorable conformation of the helix 44.