Structure and expression of the cDNA for the C isozyme of phosphofructo-1-kinase from rabbit brain.

摘要: Abstract Rabbit brain contains three phosphofructo-1-kinase (PFK) isozymic subunits designated A, B, and C. The primary structures of the first these two isozyme types have been determined previously. C rabbit was isolated by immunoaffinity chromatography subjected to proteolytic chemical digestion. A large number peptides were sequenced, total amino acids identified being equal about 80% structure. sequence cDNA derived from mRNA for polymerase chain reaction fragments synthesized using oligonucleotides designed on basis peptide sequences. deduced size 86,371 Da, slightly larger than PFKs described acid identity with 68.9% a range other sequenced mammalian 67-69%. Using data plus previously published modification, assignments 6 organic ligand binding sites PFK inferred. full-length cloned into expressed in Escherichia coli. Phosphofructokinase purified homogeneity bacterial extracts.