Prolyl aminopeptidase is also present in Enterobacteriaceae: cloning and sequencing of the Hafnia alvei enzyme-gene and characterization of the expressed enzyme.
The Hafnia alvei prolyl aminopeptidase gene (hpap) was cloned and sequenced, the expressed enzyme (HPAP) was purified to homogeneity and thoroughly characterized. An open reading frame of 1,281 bp was found to code for the enzyme, resulting in a protein of 427 amino acids with a molecular weight of 48,577. HPAP resembles the Aeromonas sobria enzyme, having 45% identity and the same distinctive properties with respect to size and substrate specificities. Both enzyme show similar chromatographic behavior, and HPAP could be purified following the procedure previously described for the Aeromonas enzyme. HPAP was found to be resistant to diisopropylphosphofluoridate as are most of the prolyl aminopeptidases hitherto described. In spite of this similarity, no inhibition by 1 mM p-chloromercuribenzoate solution could be detected. Significant inhibition was, however, observed when the enzyme was incubated with 3,4-dichloroisocoumarin. This study confirms the presence of two types of prolyl aminopeptidases, of which the Hafnia and Aeromonas enzymes constitute one group and the Bacillus, Neisseria, and Lactobacillus enzymes the other, and describes the cloning of the first prolyl aminopeptidase gene from an Enterobacteriaceae.
Kitazono
A
A
School of Pharmaceutical Sciences, Nagasaki University.
Kitano
A
A
Kabashima
T
T
Ito
K
K
Yoshimoto
T
T
D000333:Aeromonas / Q000201:enzymology; D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000235:genetics* / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology; D001483:Base Sequence; D002729:Chloromercuribenzoates / Q000494:pharmacology; D003001:Cloning, Molecular; D003374:Coumarins / Q000494:pharmacology; D004269:DNA, Bacterial; D004591:Electrophoresis, Polyacrylamide Gel; D004755:Enterobacteriaceae / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D049934:Isocoumarins; D007778:Lactobacillus / Q000201:enzymology; D008969:Molecular Sequence Data; D009343:Neisseria / Q000201:enzymology; D016366:Open Reading Frames / Q000235:genetics; D015183:Restriction Mapping; D013379:Substrate Specificity; D020245:p-Chloromercuribenzoic Acid
D002729:Chloromercuribenzoates; D003374:Coumarins; D004269:DNA, Bacterial; D049934:Isocoumarins; C046235:3,4-dichloroisocoumarin; D020245:p-Chloromercuribenzoic Acid; D000626:Aminopeptidases; C022450:prolyl aminopeptidase