Modulation of the catalytic rate of Cu,Zn superoxide dismutase in single and double mutants of conserved positively and negatively charged residues

Fabio Polticelli, Grazia Bottaro, Andrea Battistoni, Maria Teresa Carrí, Kristina Djinovic-Carugo, Martino Bolognesi, Peter O'Neill, Giuseppe Rotilio, Alessandro Desideri

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

The catalytic rate of four single and three double mutants of Xenopus laevis Cu,Zn superoxide dismutase B, neutralized at Lys120, Asp130, Glu131, and Lys134, has been determined by pulse radiolysis as a function of ionic strength. Neutralization of Glu131 increases the catalytic rate by 80% at low ionic strength, but the effect is reduced to 50% at physiological ionic strength. The rate is unperturbed upon neutralization of Asp130, while neutralization of either of the two lysines drastically decreases the enzyme activity. The Lys120Leu-Lys134Thr and Lys134Thr-Asp130Gln double mutations have an additive and a compensative effect, respectively, on the activity values, while neutralization of the Glu131-Lys134 pair, which also has a compensative effect, gives rise to a faster enzyme at any ionic strength value. The effects observed in the single Asp130Gln and Lys120Leu mutants differ from those reported on human or bovine enzymes [Getzoff et al. (1992) Nature (London) 358, 347-351; Sines et al. (1990) Biochemistry 29, 9403-9412], indicating that some residues occupying the same position in the linear sequence of different Cu,Zn superoxide dismutases have a different functional weight. Our results also suggest that the strategy of multiple charge mutation may be a promising approach in order to increase the catalytic rate of Cu,Zn SODs independently of ionic strength.

Original languageEnglish
Pages (from-to)6043-6049
Number of pages7
JournalBiochemistry
Volume34
Issue number18
Publication statusPublished - 1995

Fingerprint

Ionic strength
Osmolar Concentration
Modulation
Enzymes
Pulse Radiolysis
Radiolysis
Mutation
Biochemistry
Xenopus laevis
Enzyme activity
Lysine
Superoxide Dismutase
Superoxide Dismutase-1
Weights and Measures

ASJC Scopus subject areas

  • Biochemistry

Cite this

Polticelli, F., Bottaro, G., Battistoni, A., Carrí, M. T., Djinovic-Carugo, K., Bolognesi, M., ... Desideri, A. (1995). Modulation of the catalytic rate of Cu,Zn superoxide dismutase in single and double mutants of conserved positively and negatively charged residues. Biochemistry, 34(18), 6043-6049.

Modulation of the catalytic rate of Cu,Zn superoxide dismutase in single and double mutants of conserved positively and negatively charged residues. / Polticelli, Fabio; Bottaro, Grazia; Battistoni, Andrea; Carrí, Maria Teresa; Djinovic-Carugo, Kristina; Bolognesi, Martino; O'Neill, Peter; Rotilio, Giuseppe; Desideri, Alessandro.

In: Biochemistry, Vol. 34, No. 18, 1995, p. 6043-6049.

Research output: Contribution to journalArticle

Polticelli, F, Bottaro, G, Battistoni, A, Carrí, MT, Djinovic-Carugo, K, Bolognesi, M, O'Neill, P, Rotilio, G & Desideri, A 1995, 'Modulation of the catalytic rate of Cu,Zn superoxide dismutase in single and double mutants of conserved positively and negatively charged residues', Biochemistry, vol. 34, no. 18, pp. 6043-6049.
Polticelli, Fabio ; Bottaro, Grazia ; Battistoni, Andrea ; Carrí, Maria Teresa ; Djinovic-Carugo, Kristina ; Bolognesi, Martino ; O'Neill, Peter ; Rotilio, Giuseppe ; Desideri, Alessandro. / Modulation of the catalytic rate of Cu,Zn superoxide dismutase in single and double mutants of conserved positively and negatively charged residues. In: Biochemistry. 1995 ; Vol. 34, No. 18. pp. 6043-6049.
@article{c589627c0fc746638155ec19f63f29b5,
title = "Modulation of the catalytic rate of Cu,Zn superoxide dismutase in single and double mutants of conserved positively and negatively charged residues",
abstract = "The catalytic rate of four single and three double mutants of Xenopus laevis Cu,Zn superoxide dismutase B, neutralized at Lys120, Asp130, Glu131, and Lys134, has been determined by pulse radiolysis as a function of ionic strength. Neutralization of Glu131 increases the catalytic rate by 80{\%} at low ionic strength, but the effect is reduced to 50{\%} at physiological ionic strength. The rate is unperturbed upon neutralization of Asp130, while neutralization of either of the two lysines drastically decreases the enzyme activity. The Lys120Leu-Lys134Thr and Lys134Thr-Asp130Gln double mutations have an additive and a compensative effect, respectively, on the activity values, while neutralization of the Glu131-Lys134 pair, which also has a compensative effect, gives rise to a faster enzyme at any ionic strength value. The effects observed in the single Asp130Gln and Lys120Leu mutants differ from those reported on human or bovine enzymes [Getzoff et al. (1992) Nature (London) 358, 347-351; Sines et al. (1990) Biochemistry 29, 9403-9412], indicating that some residues occupying the same position in the linear sequence of different Cu,Zn superoxide dismutases have a different functional weight. Our results also suggest that the strategy of multiple charge mutation may be a promising approach in order to increase the catalytic rate of Cu,Zn SODs independently of ionic strength.",
author = "Fabio Polticelli and Grazia Bottaro and Andrea Battistoni and Carr{\'i}, {Maria Teresa} and Kristina Djinovic-Carugo and Martino Bolognesi and Peter O'Neill and Giuseppe Rotilio and Alessandro Desideri",
year = "1995",
language = "English",
volume = "34",
pages = "6043--6049",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "18",

}

TY - JOUR

T1 - Modulation of the catalytic rate of Cu,Zn superoxide dismutase in single and double mutants of conserved positively and negatively charged residues

AU - Polticelli, Fabio

AU - Bottaro, Grazia

AU - Battistoni, Andrea

AU - Carrí, Maria Teresa

AU - Djinovic-Carugo, Kristina

AU - Bolognesi, Martino

AU - O'Neill, Peter

AU - Rotilio, Giuseppe

AU - Desideri, Alessandro

PY - 1995

Y1 - 1995

N2 - The catalytic rate of four single and three double mutants of Xenopus laevis Cu,Zn superoxide dismutase B, neutralized at Lys120, Asp130, Glu131, and Lys134, has been determined by pulse radiolysis as a function of ionic strength. Neutralization of Glu131 increases the catalytic rate by 80% at low ionic strength, but the effect is reduced to 50% at physiological ionic strength. The rate is unperturbed upon neutralization of Asp130, while neutralization of either of the two lysines drastically decreases the enzyme activity. The Lys120Leu-Lys134Thr and Lys134Thr-Asp130Gln double mutations have an additive and a compensative effect, respectively, on the activity values, while neutralization of the Glu131-Lys134 pair, which also has a compensative effect, gives rise to a faster enzyme at any ionic strength value. The effects observed in the single Asp130Gln and Lys120Leu mutants differ from those reported on human or bovine enzymes [Getzoff et al. (1992) Nature (London) 358, 347-351; Sines et al. (1990) Biochemistry 29, 9403-9412], indicating that some residues occupying the same position in the linear sequence of different Cu,Zn superoxide dismutases have a different functional weight. Our results also suggest that the strategy of multiple charge mutation may be a promising approach in order to increase the catalytic rate of Cu,Zn SODs independently of ionic strength.

AB - The catalytic rate of four single and three double mutants of Xenopus laevis Cu,Zn superoxide dismutase B, neutralized at Lys120, Asp130, Glu131, and Lys134, has been determined by pulse radiolysis as a function of ionic strength. Neutralization of Glu131 increases the catalytic rate by 80% at low ionic strength, but the effect is reduced to 50% at physiological ionic strength. The rate is unperturbed upon neutralization of Asp130, while neutralization of either of the two lysines drastically decreases the enzyme activity. The Lys120Leu-Lys134Thr and Lys134Thr-Asp130Gln double mutations have an additive and a compensative effect, respectively, on the activity values, while neutralization of the Glu131-Lys134 pair, which also has a compensative effect, gives rise to a faster enzyme at any ionic strength value. The effects observed in the single Asp130Gln and Lys120Leu mutants differ from those reported on human or bovine enzymes [Getzoff et al. (1992) Nature (London) 358, 347-351; Sines et al. (1990) Biochemistry 29, 9403-9412], indicating that some residues occupying the same position in the linear sequence of different Cu,Zn superoxide dismutases have a different functional weight. Our results also suggest that the strategy of multiple charge mutation may be a promising approach in order to increase the catalytic rate of Cu,Zn SODs independently of ionic strength.

UR - http://www.scopus.com/inward/record.url?scp=0029009273&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029009273&partnerID=8YFLogxK

M3 - Article

VL - 34

SP - 6043

EP - 6049

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 18

ER -