Effect of single-point mutations Phe41-->His and Phe143-->Glu on folding and catalytic properties of recombinant horseradish peroxidase expressed in E. coli.
Wild-type recombinant and Phe41-->His and Phe143-->Glu mutant forms of horseradish peroxidase have been expressed in E. coli and reactivated from inclusion bodies with a yield of about 25%. The purified homogeneous preparations have been studied in the reaction of ABTS oxidation. The effect of mutations on heme entrapment and kinetics of ABTS oxidation demonstrates the essential role of the replaced residues in providing the hydrophobic crevice for the non-covalent heme binding.
Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Russian Federation.
I G
IG
Gazaryan
V V
VV
Doseeva
A G
AG
Galkin
V I
VI
Tishkov
D001483:Base Sequence; D001665:Binding Sites; D002384:Catalysis; D004926:Escherichia coli / Q000235:genetics; D018698:Glutamic Acid / Q000235:genetics; D006418:Heme / Q000378:metabolism; D006639:Histidine / Q000235:genetics; D006735:Horseradish Peroxidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010649:Phenylalanine / Q000235:genetics; D017354:Point Mutation; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship
D011994:Recombinant Proteins; D018698:Glutamic Acid; D006418:Heme; D010649:Phenylalanine; D006639:Histidine; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase