title: Construction, expression, and preliminary characterization of chimeric class mu glutathione S-transferases with altered catalytic properties.
abstract: An expression plasmid for isoenzyme 3-3 of rat liver glutathione S-transferase has been constructed from the cDNA clone pGTA/C44 and the pAS expression vector pMG27NS, and used for the efficient production of the enzyme in the Escherichia coli strain M5219. The plasmid has also been manipulated, through the use of synthetic linkers, to encode chimeric polypeptides in which short sequences of the closely related isoenzyme 4-4 have been substituted into the N-terminal and C-terminal variable domains of isoenzyme 3-3. The chimeric polypeptides designated 4(9)3(208), 3(209)4(8), and 4(9)3(200)4(8) are expressed with varying degrees of efficiency in E. coli. The active dimeric holoenzymes 3-3, (4(9)3(208]2, (3(209)4(8]2, and (4(9)3(200)4(8]2 can be isolated. The spectroscopic and kinetic properties of the chimeric enzymes are significantly different than the native enzyme.
authors.affiliation: Department of Chemistry and Biochemistry, University of Maryland, College Park 20742.
authors.forename: P H
authors.initials: PH
authors.lastname: Zhang
authors.forename: R N
authors.initials: RN
authors.lastname: Armstrong
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002678:Chimera; D002942:Circular Dichroism; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data
chemical_list: D007527:Isoenzymes; D005982:Glutathione Transferase