Cloning and expression of a catalytic core bovine brain 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase.
A cDNA encoding the catalytic core of a novel brain 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase isoenzyme was isolated from a lambda gt10 bovine brain library. This brain cDNA begins and ends in an open reading frame encoding a peptide of 476 amino acids. This peptide contains both the catalytic kinase and bisphosphatase domains and has an overall 65% and 67% indentity with the bovine heart and liver isozymes, respectively, whereas the NH2 and COOH-termini are divergent. An active catalytic core brain bifunctional enzyme was expressed in E. coli using a T7 RNA polymerase-based expression system. These results support the presence of a distinct gene coding for the protein in bovine brain.
Ventura
F
F
Unitat de Bioquimica, Universitat de Barcelona, Spain.
Ambrosio
S
S
Bartrons
R
R
el-Maghrabi
M R
MR
Lange
A J
AJ
Pilkis
S J
SJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D003001:Cloning, Molecular; D017931:DNA Primers; D004790:Enzyme Induction; D004926:Escherichia coli; D015870:Gene Expression; D015723:Gene Library; D007527:Isoenzymes / Q000096:biosynthesis / Q000737:chemistry; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008297:Male; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000096:biosynthesis* / Q000737:chemistry*; D009206:Myocardium / Q000201:enzymology; D009928:Organ Specificity; D025481:Phosphofructokinase-2; D010744:Phosphoric Monoester Hydrolases / Q000096:biosynthesis* / Q000737:chemistry*; D010770:Phosphotransferases / Q000096:biosynthesis* / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013737:Testis / Q000201:enzymology
D017931:DNA Primers; D007527:Isoenzymes; D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D010770:Phosphotransferases; D025481:Phosphofructokinase-2; D010744:Phosphoric Monoester Hydrolases