Identification and characterization of prokaryotic dipeptidyl-peptidase 5 from Porphyromonas gingivalis.
Porphyromonas gingivalis, a Gram-negative asaccharolytic anaerobe, is a major causative organism of chronic periodontitis. Because the bacterium utilizes amino acids as energy and carbon sources and incorporates them mainly as dipeptides, a wide variety of dipeptide production processes mediated by dipeptidyl-peptidases (DPPs) should be beneficial for the organism. In the present study, we identified the fourth P. gingivalis enzyme, DPP5. In a dpp4-7-11-disrupted P. gingivalis ATCC 33277, a DPP7-like activity still remained. PGN_0756 possessed an activity indistinguishable from that of the mutant, and was identified as a bacterial orthologue of fungal DPP5, because of its substrate specificity and 28.5% amino acid sequence identity with an Aspergillus fumigatus entity. P. gingivalis DPP5 was composed of 684 amino acids with a molecular mass of 77,453, and existed as a dimer while migrating at 66 kDa on SDS-PAGE. It preferred Ala and hydrophobic residues, had no activity toward Pro at the P1 position, and no preference for hydrophobic P2 residues, showed an optimal pH of 6.7 in the presence of NaCl, demonstrated Km and kcat/Km values for Lys-Ala-MCA of 688 μM and 11.02 μM(-1) s(-1), respectively, and was localized in the periplasm. DPP5 elaborately complemented DPP7 in liberation of dipeptides with hydrophobic P1 residues. Examinations of DPP- and gingipain gene-disrupted mutants indicated that DPP4, DPP5, DPP7, and DPP11 together with Arg- and Lys-gingipains cooperatively liberate most dipeptides from nutrient oligopeptides. This is the first study to report that DPP5 is expressed not only in eukaryotes, but also widely distributed in bacteria and archaea.
['Ohara-Nemoto|Yuko|Y|', 'Rouf|Shakh M A|SM|', 'Naito|Mariko|M|', 'Yanase|Amie|A|', 'Tetsuo|Fumi|F|', 'Ono|Toshio|T|', 'Kobayakawa|Takeshi|T|', 'Shimoyama|Yu|Y|', 'Kimura|Shigenobu|S|', 'Nakayama|Koji|K|', 'Saiki|Keitarou|K|', 'Konishi|Kiyoshi|K|', 'Nemoto|Takayuki K|TK|']
D001232:Aspergillus fumigatus D002384:Catalysis D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases D005656:Fungal Proteins D019897:Periplasm D033903:Periplasmic Proteins D016966:Porphyromonas gingivalis D017386:Sequence Homology, Amino Acid
Amino Acid Metabolism Peptidases Periodontal Disease Subcellular Fractionation