The role of electrostatic factors in the enzyme-substrate encounter process of manganese and iron containing superoxide dismutases has been studied in the enzyme from Propionibacterium shermanii by chemical neutralization of lysine residues and site-directed mutagenesis of the highly conserved residue Lys175. Lysine residues have been neutralized by carbamoylation and Lys175 has been selectively replaced by isoleucine and arginine, Catalytic measurements show a dramatic decrease of the activity in the chemically modified enzyme, Electrostatic potential calculations evidence in the modified enzyme a large contraction of the positive potential areas which surround the active sites in the-native enzyme, indicating that electrostatic factors are critical in the enzyme-substrate encounter process of Mn- and Fe-superoxide dismutases. The activity drastically decreases also in Lys175-->Ile but not in the Lys175-->Arg mutant. Brownian dynamics simulations indicate that the decrease of activity in the Lys175-->Ile mutant cannot be due only to a decrease of the enzyme-substrate association rate, suggesting that Lys175 plays a relevant role also in the structural stabilization of the active site.

Gabbianelli, R., Battistoni, A., Polticelli, F., Meier, B., Schmidt, M., Rotilio, G., et al. (1997). Effect of Lys175 mutation on structure function properties of Propionibacterium shermanii superoxide dismutase. PROTEIN ENGINEERING, 10(9), 1067-1070.

Effect of Lys175 mutation on structure function properties of Propionibacterium shermanii superoxide dismutase

BATTISTONI, ANDREA;ROTILIO, GIUSEPPE;DESIDERI, ALESSANDRO
1997-01-01

Abstract

The role of electrostatic factors in the enzyme-substrate encounter process of manganese and iron containing superoxide dismutases has been studied in the enzyme from Propionibacterium shermanii by chemical neutralization of lysine residues and site-directed mutagenesis of the highly conserved residue Lys175. Lysine residues have been neutralized by carbamoylation and Lys175 has been selectively replaced by isoleucine and arginine, Catalytic measurements show a dramatic decrease of the activity in the chemically modified enzyme, Electrostatic potential calculations evidence in the modified enzyme a large contraction of the positive potential areas which surround the active sites in the-native enzyme, indicating that electrostatic factors are critical in the enzyme-substrate encounter process of Mn- and Fe-superoxide dismutases. The activity drastically decreases also in Lys175-->Ile but not in the Lys175-->Arg mutant. Brownian dynamics simulations indicate that the decrease of activity in the Lys175-->Ile mutant cannot be due only to a decrease of the enzyme-substrate association rate, suggesting that Lys175 plays a relevant role also in the structural stabilization of the active site.
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Rilevanza internazionale
Articolo
Sì, ma tipo non specificato
Settore BIO/10
English
Con Impact Factor ISI
superoxide dismutase; site-directed mutagenesis; electrostatic interactions; Poisson-Boltzmann; Brownian dynamic
Gabbianelli, R., Battistoni, A., Polticelli, F., Meier, B., Schmidt, M., Rotilio, G., et al. (1997). Effect of Lys175 mutation on structure function properties of Propionibacterium shermanii superoxide dismutase. PROTEIN ENGINEERING, 10(9), 1067-1070.
Gabbianelli, R; Battistoni, A; Polticelli, F; Meier, B; Schmidt, M; Rotilio, G; Desideri, A
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/55169
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