Cited by 399Metal-Catalyzed Oxygenation MechanismsProtein Structure and DynamicsEnzyme Structure and Function

Faster superoxide dismutase mutants designed by enhancing electrostatic guidance

Getzoff, Elizabeth D., Cabelli, Diane E., Fisher, Cindy L., Parge, Hans E., Viezzoli, Maria Silvia, Banci, Lucia, Hallewell, Robert A.·1992·Nature

Abstract

The enzyme Cu, Zn superoxide dismutase (SOD) protects against oxidative damage by dismuting the superoxide radical O2-. to molecular oxygen and hydrogen peroxide at the active-site Cu ion in a reaction that is rate-limited by diffusion and enhanced by electrostatic guidance. SOD has evolved to be one of the fastest enzymes known (V(max) approximately 2 x 10(9) M-1 s-1). The new crystal structures of human SOD show that amino-acid site chains that are implicated in electrostatic guidance (Glu 132, Glu 133 and Lys 136) form a hydrogen-bonding network. Here we show that site-specific mutants that increase local positive charge while maintaining this orienting network (Glu----Gln) have faster reaction rates and increased ionic-strength dependence, matching brownian dynamics simulations incorporating electrostatic terms. Increased positive charge alone is insufficient: one charge reversal (Glu----Lys) mutant is slower than the equivalent charge neutralization (Glu----Gln) mutant, showing that the newly introduced positive charge disrupts the orienting network. Thus, electrostatically facilitated diffusion rates can be increased by design, provided the detailed structural integrity of the active-site electrostatic network is maintained.

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Crossref · 10.1038/358347a0

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  • Several studies have been carried out using pulsed radiolysis to generate HO2•/O2•− and to understand the Cu,Zn-SOD-catalysed reaction of superoxide dismutation.

    “Several studies have been carried out using pulsed radiolysis to generate HO2•/O2•− and to understand the Cu,Zn-SOD-catalysed reaction of superoxide dismutation [74,75,76,77], Figure 32.”

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Authors
Getzoff, E. D., Cabelli, D. E., Fisher, C. L., Parge, H. E., Viezzoli, M. S., Banci, L., & Hallewell, R. A.
Affiliations
  • Scripps Research Institute (US)
  • Brookhaven National Laboratory (US)
  • University of Florence (IT)
Venue
Nature
Published
1992-07
Volume
358
Issue
6384
Pages
347-351
ISSN
  • 0028-0836 (ISSN-L)
  • 1476-4687
OpenAlex
W2051555156
Cited by
399
References
30
Citations by year
2026: 5 · 2025: 9 · 2024: 5 · 2023: 7 · 2022: 7 · 2021: 9 · 2020: 11 · 2019: 11 · 2018: 9 · 2017: 13 · 2016: 7 · 2015: 9 · 2014: 11 · 2013: 7 · 2012: 9
FWCI
26.78
Type
journal article
Language
en
Open access
closed
Locations
  • Nature (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Physical Sciences › Chemistry › Inorganic Chemistry › Metal-Catalyzed Oxygenation Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
Keywords
superoxide dismutase, enzyme kinetics, site-directed mutagenesis, hydrogen bond network, charge reversal, ionic strength dependence, diffusion rate, Brownian dynamics
MeSH terms
Amino Acid Sequence, Computer Simulation, Electrochemistry, Glutamates, Humans, Hydrogen Bonding, Hydrogen-Ion Concentration, Kinetics, Lysine, Models, Molecular, Osmolar Concentration, Protein Conformation, Superoxide Dismutase, Mutagenesis, Site-Directed, Glutamic Acid

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