[Protein engineering of uridine phosphorylase from Escherichia coli K-12. II. Comparative study of hybrid and mutant forms of uridine phosphorylases].
Genes for hybrid uridine phosphorylases (UPases) consisting of fragments of amino acid sequences of UPases from Escherichia coli and Salmonella typhimurium were constructed. Producing strains of the corresponding proteins were genetically engineered. Mutant forms of the E. coli K-12 UPase were produced by site-directed mutagenesis. A comparative study of the enzyme properties of the mutant and hybrid forms of bacterial UPases was performed. It was shown that Asp27 unlike Asp5 and Asp29 residues of the E. coli UPase forms part of the active site of the protein. A scheme of the involvement of Asp27 in the binding of inorganic phosphate is proposed.
Chebotarev
D V
DV
State Research Institute of Genetics and Selection of Industrial Microorganisms, Pervyi Dorozhnyi pr. 1, Moscow, 113545 Russia.
Gul'ko
L B
LB
Veĭko
V P
VP
D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000201:enzymology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D016415:Sequence Alignment; D014543:Uridine Phosphorylase / Q000032:analysis* / Q000235:genetics
D011993:Recombinant Fusion Proteins; D014543:Uridine Phosphorylase