Design, creation, and characterization of a stable,monomeric triosephosphate isomerase.
Open Access
- 15 February 1994
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 91 (4), 1515-1518
- https://doi.org/10.1073/pnas.91.4.1515
Abstract
Protein engineering on trypanosomal triosephosphate isomerase (TIM) converted this oligomeric enzyme into a stable, monomeric protein that is enzymatically active. Wild-type TIM consists of two identical subunits that form a very tight dimer involving interactions of 32 residues of each subunit. By replacing 15 residues of the major interface loop by another 8-residue fragment, a variant was constructed that is a stable and monomeric protein with TIM activity. The length, sequence, and conformation of the designed fragment were suggested by extensive modeling.Keywords
This publication has 24 references indexed in Scilit:
- The crystal structure of an engineered monomeric triosephosphate isomerase, monoTIM: the correct modelling of an eight-residue loopStructure, 1993
- Replacing the (βα)-unit 8 of E.coli TIM with its chicken homologue leads to a stable and active hybrid enzymeProtein Engineering, Design and Selection, 1993
- Optimal protocol and trajectory visualization for conformational searches of peptides and proteinsJournal of Molecular Biology, 1992
- Comparison of the refined crystal structures of liganded and unliganded chicken, yeast and trypanosomal triosephosphate isomeraseJournal of Molecular Biology, 1992
- Atomic solvation parameters applied to molecular dynamics of proteins in solutionProtein Science, 1992
- Refined 1.83 Å structure of trypanosomal triosephosphate isomerase crystallized in the presence of 2.4 m-ammonium sulphate: A comparison with the structure of the trypanosomal triosephosphate isomerase-glycerol-3-phosphate complexJournal of Molecular Biology, 1991
- Probing the catalytic sites of triosephosphate isomerase by 31P‐NMR with reversibly and irreversibly binding substrate analoguesEuropean Journal of Biochemistry, 1991
- Surface, subunit interfaces and interior of oligomeric proteinsJournal of Molecular Biology, 1988
- Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genesJournal of Molecular Biology, 1986
- Reversible dissociation of dimeric tyrosyl-tRNA synthetase by mutagenesis at the subunit interfaceBiochemistry, 1985