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 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.
Borchert
T V
TV
European Molecular Biology Laboratory, Heidelberg, Germany.
Abagyan
R
R
Jaenicke
R
R
Wierenga
R K
RK
D000595:Amino Acid Sequence; D000818:Animals; D002942:Circular Dichroism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D015202:Protein Engineering; D017384:Sequence Deletion; D014305:Triose-Phosphate Isomerase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D014346:Trypanosoma brucei brucei / Q000201:enzymology*
D014305:Triose-Phosphate Isomerase