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Construction of a catalytically active iron superoxide dismutase by rational protein design.

Publication ,  Journal Article
Pinto, AL; Hellinga, HW; Caradonna, JP
Published in: Proc Natl Acad Sci U S A
May 27, 1997

The rational protein design algorithm DEZYMER was used to introduce the active site of nonheme iron superoxide dismutase (SOD) into the hydrophobic interior of the host protein, Escherichia coli thioredoxin (Trx), a protein that does not naturally contain a transition metal-binding site. Reconstitution of the designed protein, Trx-SOD, showed the incorporation of one high-affinity metal-binding site. The electronic spectra of the holoprotein and its N3- and F- adducts are analogous to those previously reported for native {Fe3+}SOD. Activity assays showed that {Fe3+}Trx-SOD is capable of catalyzing the dismutation of the superoxide anion; comparative studies with the unrelated wild-type E. coli iron SOD indicated that {Fe3+}Trx-SOD catalyzes the dismutation reaction at a rate on the order of 10(5) M-1s -1. The ability to design catalytically competent metalloenzymes allows for the systematic investigation of fundamental mechanistic questions concerning catalysis at transition metal centers.

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Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

May 27, 1997

Volume

94

Issue

11

Start / End Page

5562 / 5567

Location

United States

Related Subject Headings

  • Thioredoxins
  • Superoxide Dismutase
  • Recombinant Fusion Proteins
  • Protein Structure, Secondary
  • Protein Engineering
  • Mutagenesis, Site-Directed
  • Models, Structural
  • Iron
  • Escherichia coli
  • Computer Simulation
 

Citation

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Pinto, A. L., Hellinga, H. W., & Caradonna, J. P. (1997). Construction of a catalytically active iron superoxide dismutase by rational protein design. Proc Natl Acad Sci U S A, 94(11), 5562–5567. https://doi.org/10.1073/pnas.94.11.5562
Pinto, A. L., H. W. Hellinga, and J. P. Caradonna. “Construction of a catalytically active iron superoxide dismutase by rational protein design.Proc Natl Acad Sci U S A 94, no. 11 (May 27, 1997): 5562–67. https://doi.org/10.1073/pnas.94.11.5562.
Pinto AL, Hellinga HW, Caradonna JP. Construction of a catalytically active iron superoxide dismutase by rational protein design. Proc Natl Acad Sci U S A. 1997 May 27;94(11):5562–7.
Pinto, A. L., et al. “Construction of a catalytically active iron superoxide dismutase by rational protein design.Proc Natl Acad Sci U S A, vol. 94, no. 11, May 1997, pp. 5562–67. Pubmed, doi:10.1073/pnas.94.11.5562.
Pinto AL, Hellinga HW, Caradonna JP. Construction of a catalytically active iron superoxide dismutase by rational protein design. Proc Natl Acad Sci U S A. 1997 May 27;94(11):5562–5567.
Journal cover image

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

May 27, 1997

Volume

94

Issue

11

Start / End Page

5562 / 5567

Location

United States

Related Subject Headings

  • Thioredoxins
  • Superoxide Dismutase
  • Recombinant Fusion Proteins
  • Protein Structure, Secondary
  • Protein Engineering
  • Mutagenesis, Site-Directed
  • Models, Structural
  • Iron
  • Escherichia coli
  • Computer Simulation