Abstract
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.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Animals
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Base Sequence
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Binding Sites
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Brain / enzymology*
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Cattle
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Cloning, Molecular
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DNA Primers
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Enzyme Induction
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Escherichia coli
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Gene Expression
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Gene Library
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Isoenzymes / biosynthesis
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Isoenzymes / chemistry
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Kinetics
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Liver / enzymology
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Male
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Molecular Sequence Data
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Multienzyme Complexes / biosynthesis*
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Multienzyme Complexes / chemistry*
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Myocardium / enzymology
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Organ Specificity
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Phosphofructokinase-2
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Phosphoric Monoester Hydrolases / biosynthesis*
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Phosphoric Monoester Hydrolases / chemistry*
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Phosphotransferases / biosynthesis*
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Phosphotransferases / chemistry*
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Polymerase Chain Reaction
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Sequence Homology, Amino Acid
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Testis / enzymology
Substances
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DNA Primers
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Isoenzymes
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Multienzyme Complexes
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Recombinant Proteins
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Phosphotransferases
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Phosphofructokinase-2
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Phosphoric Monoester Hydrolases