title: Isolation of the leucine aminopeptidase gene from Aeromonas proteolytica. Evidence for an enzyme precursor.
abstract: The leucine aminopeptidase of Aeromonas proteolytica (EC 3.4.11.10) is a monomeric metalloenzyme having the capacity to bind two Zn2+ atoms in the active site. Structural information of this relatively small aminopeptidase that could illuminate the catalytic mechanism of the metal ions is lacking; hence, we have obtained sequences from the purified enzyme, cloned the corresponding gene, and expressed the recombinant protein in Escherichia coli. The deduced primary amino acid sequence of this secreted protease suggests a potential signal peptide at the NH2 terminus. Expression of the recombinant and native proteins in E. coli and in extracts of culture media of A. proteolytica indicates that the aminopeptidase is secreted as an active and thermosensitive 43-kDa protein that is rapidly transformed to thermostable forms of 30 and 32 kDa. Comparison of the deduced amino acid sequence of the A. proteolytica leucine aminopeptidase with other Zn(2+)-binding metalloenzymes failed to show homologies to the consensus binding sequence His-Glu-X-X-His for the metal ion.
authors.affiliation: Marion Merrell Dow Research Institute, Strasbourg, France.
authors.forename: C
authors.initials: C
authors.lastname: Guenet
authors.forename: P
authors.initials: P
authors.lastname: Lepage
authors.forename: B A
authors.initials: BA
authors.lastname: Harris
mesh_terms: D000333:Aeromonas / Q000201:enzymology*; D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D004269:DNA, Bacterial / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D007931:Leucyl Aminopeptidase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D004269:DNA, Bacterial; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D007931:Leucyl Aminopeptidase