Design, Creation, and Characterization of a Stable, Monomeric Triosephosphate Isomerase

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...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1994-02, Vol.91 (4), p.1515-1518
Main Authors: Borchert, Torben V., Abagyan, Ruben, Jaenicke, Rainer, Wierenga, Rik K.
Format: Article
Language:English
Subjects:
Active sites
Amino Acid Sequence
Amino acids
Animals
Atoms
Biochemistry
Biological and medical sciences
Biotechnology
Circular Dichroism
Design engineering
Dimers
Enzyme Stability
Enzymes
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
Gels
Methods. Procedures. Technologies
Models, Molecular
Molecular Sequence Data
Monomers
Mutagenesis, Site-Directed
Protein Conformation
Protein Engineering
Proteins
Sequence Deletion
Triose-Phosphate Isomerase - chemistry
Triose-Phosphate Isomerase - genetics
Triose-Phosphate Isomerase - metabolism
Trypanosoma
Trypanosoma brucei brucei - enzymology
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Summary: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.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.91.4.1515