# P-site
The **P-site** (peptidyl site) is one of the three transfer RNA binding sites of the ribosome, the site that holds the peptidyl-tRNA, the tRNA molecule carrying the growing polypeptide chain. Initial models of translation accounted for protein synthesis through two tRNA binding sites defined operationally: the P-site, holding the donor substrate (peptidyl-tRNA), and the A-site, holding the acceptor substrates (aminoacyl-tRNAs).[1](https://pubmed.ncbi.nlm.nih.gov/9598294/) A third site, the E-site for deacylated tRNA, was later added, giving the allosteric three-site model used today.[1](https://pubmed.ncbi.nlm.nih.gov/9598294/)
| Key fact | Detail |
|---|---|
| Definition | The ribosomal binding site for peptidyl-tRNA, the donor substrate in peptide bond formation[1](https://pubmed.ncbi.nlm.nih.gov/9598294/) |
| Initiation | The initiating methionyl-tRNA occupies the P site at the start of elongation in both prokaryotes and eukaryotes[2](https://openstax.org/books/biology-2e/pages/15-5-ribosomes-and-protein-synthesis) |
| 50S contacts | C74 and C75 of P-site tRNA form Watson-Crick base pairs with G2252 and G2251 of 23S rRNA, orienting the 3' end for peptidyl transfer[3](https://pmc.ncbi.nlm.nih.gov/articles/PMC3010847/) |
| 30S contacts | A1339 and G1338 of 16S rRNA form type I/II base triples with the P-tRNA; A1339 is critical for translation in vivo and P-site binding in vitro[3](https://pmc.ncbi.nlm.nih.gov/articles/PMC3010847/) |
| Binding strength | Peptidyl-tRNA in the P/P classical state is the strongest tRNA binding state, with high affinity for both the 30S and 50S P sites[4](https://pmc.ncbi.nlm.nih.gov/articles/PMC6581036/) |
| During elongation | After peptidyl transfer, the A-site tRNA moves into the P site and the deacylated P-site tRNA moves to the E site and is expelled[2](https://openstax.org/books/biology-2e/pages/15-5-ribosomes-and-protein-synthesis) |
## Role in the elongation cycle
Each cycle of translation elongation has three major steps: decoding, peptidyl transfer, and translocation.[3](https://pmc.ncbi.nlm.nih.gov/articles/PMC3010847/) The cycle begins with the initiator methionyl-tRNA seated in the P site.[2](https://openstax.org/books/biology-2e/pages/15-5-ribosomes-and-protein-synthesis) [Aminoacyl-tRNA](https://www.edgepedia.org/aminoacyl-trna) is delivered to the A site as part of a ternary complex with elongation factor Tu and GTP, and once it occupies the A sites of both subunits the ribosome catalyzes transfer of the peptidyl group of the P-site tRNA to the A-site aminoacyl-tRNA. This leaves peptidyl-tRNA in the A site and deacylated tRNA in the P site, the pretranslocation complex.[3](https://pmc.ncbi.nlm.nih.gov/articles/PMC3010847/)
**Translocation.** Elongation factor G then catalyzes the coupled movement of tRNA and mRNA: the peptidyl-tRNA moves from the A site into the P site, and the deacylated tRNA moves from the P site into the E site, leaving the A site vacant for the next round.[3](https://pmc.ncbi.nlm.nih.gov/articles/PMC3010847/) After peptide bond formation the A-site tRNA that now holds the growing chain moves to the P site, and the emptied P-site tRNA moves to the E site and is expelled from the ribosome.[2](https://openstax.org/books/biology-2e/pages/15-5-ribosomes-and-protein-synthesis) EF-G accelerates translocation by 4 to 5 orders of magnitude, and GTP hydrolysis stimulates the rate of translocation and EF-G turnover by about 50-fold.[3](https://pmc.ncbi.nlm.nih.gov/articles/PMC3010847/)
## Molecular contacts
The P site exists on both ribosomal subunits. In the 50S P site, C74 and C75 of the tRNA form Watson-Crick base pairs with G2252 and G2251 of 23S rRNA, orienting the 3' end of peptidyl-tRNA for peptidyl transfer. The tRNA elbow interacts with the C terminus of protein L5, and the D stem interacts with helix 69 of 23S rRNA.[3](https://pmc.ncbi.nlm.nih.gov/articles/PMC3010847/) In the 30S P site, the anticodon stem loop is contacted by proteins S9 and S13 and by 16S rRNA nucleotides including A1339 and G1338, which form type I/II base triples with base pairs in the tRNA; the loop containing these nucleotides forms a "gate" with the 790 loop between the P and E sites that appears to control tRNA movement during translocation.[3](https://pmc.ncbi.nlm.nih.gov/articles/PMC3010847/)
Binding strength and dynamics. The strongest tRNA binding state is peptidyl-tRNA in the P/P classical state, because of its high affinity for both the 30S and 50S P sites; this state enforces the minimal intersubunit rotation observed in the classical conformation.[4](https://pmc.ncbi.nlm.nih.gov/articles/PMC6581036/) Spontaneous rotations are infrequent in ribosomes carrying only peptidyl-tRNA in the P site, while pretranslocation complexes rapidly exchange between rotated and nonrotated conformations, indicating that peptidyl transfer unlocks spontaneous intersubunit rotation.[5](https://pmc.ncbi.nlm.nih.gov/articles/PMC10576065/pdf/pnas.202114979.pdf)
Early ultraviolet crosslinking of E. coli ribosomal complexes identified the proteins in contact with P-site tRNA as S7, S9, S11, L2, L4, L7/L12, and L27, showing that tRNA in the P site touches proteins of both the 30S and 50S subunits.[6](https://pubmed.ncbi.nlm.nih.gov/386278/)
## From the two-site to the three-site model
The two-site model was the minimum number of binding sites that could account for the known chemistry of protein synthesis, the attack of the amino group of aminoacyl-tRNA on the carbonyl group of peptidyl-tRNA.[7](https://febs.onlinelibrary.wiley.com/doi/10.1016/S0014-5793%2802%2902327-X) Discovery of the E-site, specific for deacylated tRNAs, produced the allosteric three-site model, whose two major features are the reciprocal relationship of A-site and E-site occupation, and simultaneous codon-anticodon interactions of both tRNAs present at the elongating ribosome.[1](https://pubmed.ncbi.nlm.nih.gov/9598294/) Allosterically, when a deacylated tRNA occupies the E site in addition to a tRNA at the P site, the A site cannot be occupied by aminoacyl-tRNA at low temperature and only poorly by the EF-Tu ternary complex.[8](https://www.sciencedirect.com/science/article/pii/S0021925818676288)
Base-specific footprinting later led to the conclusion that there are at least five distinct tRNA binding sites on the ribosome, A and P sites on the 30S subunit and A, P, and E sites on the 50S subunit; the crystal structure of the 70S ribosome bound with tRNAs confirmed a 30S E site as well, so binding sites for A, P, and E tRNAs exist on both subunits.[7](https://febs.onlinelibrary.wiley.com/doi/10.1016/S0014-5793%2802%2902327-X) The first atomic model of the 70S ribosome showed the general positions of tRNAs bound to the A, P, and E sites, and higher-resolution structures have since detailed the interactions made by tRNAs with the P and E sites.[9](https://mediatheque.lindau-nobel.org/recordings/33649/2014-from-the-structure-and-function-of-the-ribosome-to-new-antibiotics)
## References
1. [Ribosomal tRNA binding sites: three-site models of translation (PubMed)](https://pubmed.ncbi.nlm.nih.gov/9598294/)
2. [Ribosomes and Protein Synthesis, Biology 2e, OpenStax](https://openstax.org/books/biology-2e/pages/15-5-ribosomes-and-protein-synthesis)
3. [Shoji, Walker & Fredrick (2009). Ribosomal Translocation: One Step Closer to the Molecular Mechanism. ACS Chemical Biology](https://pmc.ncbi.nlm.nih.gov/articles/PMC3010847/)
4. [The Ribosome Moves: RNA Mechanics and Translocation (PMC)](https://pmc.ncbi.nlm.nih.gov/articles/PMC6581036/)
5. [The unlocking of ribosome dynamics by peptidyl transfer (PNAS)](https://pmc.ncbi.nlm.nih.gov/articles/PMC10576065/pdf/pnas.202114979.pdf)
6. [Polynucleotide-protein interactions in the translation system: proteins interacting with tRNA in the A- and P-sites of E. coli ribosomes (PubMed)](https://pubmed.ncbi.nlm.nih.gov/386278/)
7. [Noller (2002). Translocation of tRNA during protein synthesis. FEBS Letters](https://febs.onlinelibrary.wiley.com/doi/10.1016/S0014-5793%2802%2902327-X)
8. [Allosteric interactions between the ribosomal transfer RNA-binding sites A and E (JBC)](https://www.sciencedirect.com/science/article/pii/S0021925818676288)
9. [Thomas A. Steitz lecture, Lindau Mediatheque](https://mediatheque.lindau-nobel.org/recordings/33649/2014-from-the-structure-and-function-of-the-ribosome-to-new-antibiotics)
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*Topic: Encyclopedia › Life and health › Biological foundations › Cell biology › Organelles › Ribosomes and cytoplasmic translation › Elongation, termination, and release*
*Initially written Sep 17, 2026 · Reviewed: Sep 30, 2026; Oct 21, 2026 · Edited: Oct 15, 2026; Oct 21, 2026 · Last review: Oct 21, 2026*
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