Sso7d is a small basic protein consisting of 62 amino acids isolated from the thermoacidophilic archeobacterium Sulfolobus solfataricus. The protein is endowed with DNA binding properties, RNase activity, and the capability of rescuing aggregated proteins in the presence of ATP. In this study, the electrostatic properties of Sso7d are investigated by using the Poisson-Boltzmann calculation of the surface potential distribution and following by NMR spectroscopy the proton chemical shift pH titration of acidic residues. Although the details of the catalytic mechanism still have to be defined, the results from NMR experiments confirm the possible involvement of Glu35 as the proton acceptor in the catalytic reaction, as seen by its abnormally high pK(a) value. Poisson-Boltzmann calculations and NMR titration shifts suggest the presence of a possible hydrogen bond between Glu35 and Tyr33, with a consequent rather rigid arrangement at these positions. Comparison with RNase T1 suggests that Tyr7 may be a good candidate for acting as a proton donor in the active site of Sso7d as shown by its low phenolic pKa of similar to9.3. Titration experiments performed with the UpA, a RNA dinucleotide model, showed that the protein residues affected by the interaction are mainly located in a different region with respect to the surface affected by DNA recognition, in good agreement with the surface potential distribution found with electrostatic calculations.
|Citazione:||Consonni, R., Arosio, I., Belloni, B., Fogolari, F., Fusi, P., Shehi, E., et al. (2003). Investigations of Sso7d catalytic residues by NMR titration shifts and electrostatic calculations. BIOCHEMISTRY, 42(6), 1421-1429.|
|Tipo:||Articolo in rivista - Articolo scientifico|
|Carattere della pubblicazione:||Scientifica|
|Titolo:||Investigations of Sso7d catalytic residues by NMR titration shifts and electrostatic calculations|
|Autori:||Consonni, R; Arosio, I; Belloni, B; Fogolari, F; Fusi, P; Shehi, E; Zetta, L|
|Data di pubblicazione:||18-feb-2003|
|Appare nelle tipologie:||01 - Articolo su rivista|