TABLE 1.
Characteristics of the available LepA structures in complex with 70S ribosomes
| Structure information | PRE complexes |
POST complexes |
|||||
|---|---|---|---|---|---|---|---|
| LepA-GDPNP-PRE | LepA-GDPNP-PRE (major group) | LepA-GDPCP-PRE | LepA-GDP-POST | LepA-GDPNP-POST (minor group) | LepA-GDPCP-POST (major group) | LepA-GDPCP-POST (minor group) | |
| Reference | 25 | 23 | 15 | 27 | 23 | 35 | 35 |
| Method | Cryo-EM | Cryo-EM | X-ray crystallography | X-ray crystallography | Cryo-EM | Cryo-EM | Cryo-EM |
| Resolution (Å) | 10.9 | 3.2 | 2.6 | 2.9 | 3.7 | 3.8 | 5.7 |
| Organism | Escherichia coli | Escherichia coli | Thermus thermophilus | Thermus thermophilus | Escherichia coli | Thermus thermophilus | Thermus thermophilus |
| mRNA (nt) | MF-mRNAa (46), AUG-UUC | MF-mRNA (81), SDb (GGAGG) AUG-UUC | MF-mRNA (24), SD (GGAGG) AUG-UUC | mRNA (18), SD (GGAGG) UUC | MF-mRNA (81), SD (GGAGG) AUG-UUC | MF-mRNA (25), SD (GGAGG) AUG-UUC | MF-mRNA (25), SD (GGAGG) AUG-UUC |
| A-tRNA (state) | Ac[14C]Phe-tRNAPhe (A/L) | Ac[14C]Phe-tRNAPhe (A/4) | Phe-tRNAPhe (A/L) | None | None | None | None |
| P-tRNA | [32P]deacyl-tRNAfMet | Deacyl-tRNAfMet | fMet-tRNAfMet | Phe-tRNAPhe | Ac[14C]Phe-tRNAPhe | tRNAfMet | tRNAfMet |
| E-tRNA | None | None | None | tRNAPhe (modeled) | Deacyl-tRNAfMet | tRNAPhe | tRNAPhe |
| 30S rotation (°) | Unrotated | Unrotated | Unrotated | Clockwise (5) | Clockwise (4) | Unrotated | Counterclockwise (5.6) |
| PDB ID | 3DEG | 3JCE | 5J8B | 4W2E | 3JCD | 51MQ | 51MR |
| Prepn | LepA and GDPNP incubated with POST state 70S | LepA and GDPNP incubated with POST state 70S | LepA and GDPCP incubated with PRE state 70S | LepA and GDPCP incubated with 70S and P-site Phe-tRNAPhe | PRE state 70S incubated with EF-G to yield POST state; LepA and GDPNP incubated with POST state 70S | LepA and GDPCP incubated with PRE state 70S | LepA and GDPCP incubated with PRE state 70S |
| Comments | Missing last 55 aa of CTD | None | Engineered L9 fusion; highest-resolution structure so far | Engineered L9 fusion | Only modeled 26 residues of CTD | Engineered L9 fusion | Engineered L9 fusion |
a
MF-mRNA, mRNa containing the codons for methionine (M) and phenylalanine (F).
b
SD, Shine-Dalgarno element. The sequences of the SD (in parentheses) and the codons are indicated.