1. Drosophila melanogaster acylphosphatase: A common ancestor for acylphosphatase isoenzymes of vertebrate species
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Alessandro Pieri, Maria Pia Bozzetti, Giovanni Raugei, Gianfranco Liguri, Francesca Magherini, Giampietro Ramponi, Niccolò Taddei, Cristina Cecchi, Pieri, A., Magherini, F., Liguri, G., Raugei, G., Taddei, N., Bozzetti, Maria Giuseppina, Cecchi, C., and Ramponi, G.
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Molecular Sequence Data ,Biophysics ,Acylphosphatase ,Biochemistry ,Isozyme ,Gene product ,Evolution, Molecular ,Open Reading Frames ,Structural Biology ,Complementary DNA ,Genetics ,Melanogaster ,Escherichia coli ,Animals ,Humans ,Amino Acid Sequence ,Cloning, Molecular ,Molecular Biology ,Gene ,Conserved Sequence ,Phylogeny ,acylphosphatase ,biology ,Sequence Homology, Amino Acid ,Cell Biology ,biology.organism_classification ,Recombinant Proteins ,Acid Anhydride Hydrolases ,Isoenzymes ,human muscle acylphosphatase ,Open reading frame ,Kinetics ,Drosophila melanogaster ,Vertebrates ,Mutagenesis, Site-Directed ,Human muscle acylphosphatase ,Drosophila ,Sequence Alignment - Abstract
An open reading frame encoding a putative acylphosphatase was found in Drosophila melanogaster. The corresponding gene product shows 40% identity and 22 additional amino acid residues at the C-terminus as compared to muscle- and common-type human acylphosphatases. Moreover, all the residues involved in the catalytic mechanism of vertebrate enzymes are conserved in the D. melanogaster acylphosphatase. The D. melanogaster protein and a deletion mutant, similar in length to vertebrate acylphosphatases, were produced by cloning the corresponding cDNA in Escherichia coli. The wild-type enzyme is a protein with a well-established three-dimensional fold and a markedly reduced conformational stability as compared to vertebrate isoenzymes. The specific activity of the enzyme is significantly lower than that found in vertebrate enzymes though the substrate binding capability is basically unaltered. The deletion of 22 residues does not cause a significant change in kcat, while affecting the apparent binding parameters. This work suggests that the genes encoding the vertebrate enzymes originate from an ancestor gene by duplication and subsequent evolution.
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