Doucet, Nicolas 
ORCID: https://orcid.org/0000-0002-1952-9380; Jayasundera, Thusitha B.; Simonović, Miljan et Loria, J. Patrick
(2010).
The crystal structure of ribonuclease A in complex with thymidine-3'-monophosphate provides further insight into ligand binding
Proteins
, vol. 78
, nº 11.
pp. 2459-2468.
DOI: 10.1002/prot.22754.
Résumé
Thymidine-3'-monophosphate (3'-TMP) is a competitive inhibitor analogue of the 3'-CMP and 3'-UMP natural product inhibitors of bovine pancreatic ribonuclease A (RNase A). Isothermal titration calorimetry experiments show that 3'-TMP binds the enzyme with a dissociation constant (K(d)) of 15 microM making it one of the strongest binding members of the five natural bases found in nucleic acids (A, C, G, T, and U). To further investigate the molecular properties of this potent natural affinity, we have determined the crystal structure of bovine pancreatic RNase A in complex with 3'-TMP at 1.55 A resolution and we have performed NMR binding experiments with 3'-CMP and 3'-TMP. Our results show that binding of 3'-TMP is very similar to other natural and non-natural pyrimidine ligands, demonstrating that single nucleotide affinity is independent of the presence or absence of a 2'-hydroxyl on the ribose moiety of pyrimidines and suggesting that the pyrimidine binding subsite of RNase A is not a significant contributor of inhibitor discrimination. Accumulating evidence suggests that very subtle structural, chemical, and potentially motional variations contribute to ligand discrimination in this enzyme.
| Type de document: | Article |
|---|---|
| Mots-clés libres: | - |
| Centre: | Centre INRS-Institut Armand Frappier |
| Date de dépôt: | 29 juin 2024 21:46 |
| Dernière modification: | 29 juin 2024 21:46 |
| URI: | https://espace.inrs.ca/id/eprint/14610 |
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