Functional complementation of yeast phosphofructokinase mutants by the non-allosteric enzyme from Dictyostelium discoideum.
Phosphofructokinase (PFK) from yeast has been replaced by the non-allosteric isozyme from the slime mold Dictyostelium discoideum. This has been achieved by overexpression of the latter in a PFK-deficient strain of Saccharomyces cerevisiae under the control of the PFK2 promoter. Transformants complemented the glucose-negative growth phenotype exhibiting generation times on glucose-containing media similar to those of an untransformed strain being wild-type for yeast PFK genes. The PFK produced reacted with an antibody against D. discoideum PFK. It exhibited the same subunit size, quaternary structure and kinetic parameters than those of the wild-type enzyme, and was also devoid of specific regulatory properties.
Estévez
A M
AM
Departamento de Bioquímica de la UAM, Facultad de Medicina, Universidad Autónoma, Madrid, Spain.
Heinisch
J J
JJ
Aragón
J J
JJ
D000494:Allosteric Regulation; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004023:Dictyostelium / Q000201:enzymology* / Q000235:genetics; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009154:Mutation; D010732:Phosphofructokinase-1 / Q000032:analysis / Q000235:genetics*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D014170:Transformation, Genetic
D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D010732:Phosphofructokinase-1