Paper Report for: Clery-Barraud_2002_Eur.J.Biochem_269_4297
Reference
Title: Pressure and heat inactivation of recombinant human acetylcholinesterase. Importance of residue E202 for enzyme stability Clery-Barraud C, Ordentlich A, Grosfeld H, Shafferman A, Masson P Ref: European Journal of Biochemistry, 269:4297, 2002 : PubMed
The effects of pressure on structure and activity of recombinant human acetylcholinesterase (rHuAChE) were investigated up to a pressure of 300 MPa using gel electrophoresis under elevated hydrostatic pressure, fluorescence of bound 8-anilinonaphthalene-1-sulfonate (ANS) and activity measurements following exposure to high pressure. Study of wild-type enzyme and three single mutants (D74N, E202Q, E450A) and one sextuple mutant (E84Q/E292A/D349N/E358Q/E389Q/D390N) showed that pressure exerts a differential action on wild-type rHuAChE and its mutants, allowing estimation of the contribution of carboxylic amino acid side-chains to enzyme stability. Mutation of negatively charged residues D74 and E202 by polar side-chains strengthened heat or pressure stability. The mutation E450A and the sextuple mutation caused destabilization of the enzyme to pressure. Thermal inactivation data on mutants showed that all of them were stabilized against temperature. In conclusion, pressure and thermal stability of mutants provided evidence that the residue E202 is a determinant of structural and functional stability of HuAChE.
Clery-Barraud C, Ordentlich A, Grosfeld H, Shafferman A, Masson P (2002) Pressure and heat inactivation of recombinant human acetylcholinesterase. Importance of residue E202 for enzyme stability European Journal of Biochemistry269: 4297-307
Clery-Barraud C, Ordentlich A, Grosfeld H, Shafferman A, Masson P (2002) European Journal of Biochemistry269: 4297-307