Faster superoxide dismutase mutants designed by enhancing electrostatic guidance
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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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.”
From Superoxide Anion Chemistry—Its Role at the Core of the Innate Immunity (2023) · cites this paper as [77]
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Self-citation
- 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)
- ORCID
- Banci, Lucia
- 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
- Licence
- www.springer.com/tdm (tdm)
- 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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