The gene encoding dipeptidyl aminopeptidase BI from Pseudomonas sp. WO24: cloning, sequencing and expression in Escherichia coli

Ogasawara, Wataru; Kobayashi, Go; Ishimaru, Seiji; Okada, Hirofumi; Morikawa, Yasushi

doi:10.1016/s0378-1119(97)00590-8

Cited by 6 publications

(7 citation statements)

References 24 publications

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“…3) showed a clear (57% height of the origin peak) noncrystallographic twofold axis, indicating that the number of molecules in the asymmetric unit of the DAP BII crystal would be an even number. This observation is consistent with the dimeric nature of DAP BII (Ogasawara et al, 1998) and indicates that the number of molecules in the asymmetric unit of the DAP BII crystal must be two. A search for heavy-atom derivatives intended for phasing by multiple isomorphous replacement and preparation of selenomethioninesubstituted DAP BII using LeMaster medium (Hendrickson et al, 1990) Stereographic projection of the self-rotation function in spherical polar angles at = 180 for the DAP BII crystal.…”

Section: X-ray Analysessupporting

confidence: 82%

“…The hydrolytic activities of recombinant DAP BII for synthetic substrates were 3.4 AE 0.1 mmol min À1 mg À1 (U mg À1 ) and 10 AE 0.2 U mg À1 for Gly-Phe-pNA and Ala-Ala-pNA, respectively, which were compar- able to those of native DAP BII (3.3 AE 0.1 and 9.5 AE 0.2 U mg À1 for Gly-Phe-pNA and Ala-Ala-pNA, respectively). The recombinant enzyme was equally stable to the native enzyme at temperatures below 293 K for 30 min (Ogasawara et al, 1998).…”

Section: Resultsmentioning

confidence: 99%

“…Briefly, an Escherichia coli Rosetta 2(DE3)pLacI transformant harbouring the mature DAP BII sequence (Gly25-Lys722) with the signal peptide of DAP BIII from P. mexicana WO24 (Ogasawara et al, 1998) inserted into pET-22b expression plasmid was used for the production of recombinant DAP BII. The mature DAP BII was composed of 698 amino acids with a theoretical molecular mass of 76 299.4 Da and an isoelectric point of 6.30.…”

Section: Overproduction and Purificationmentioning

confidence: 99%

“…We have reported the identification, purification and characterization of DAP BI (bacterial dipeptidyl aminopeptidase; Ogasawara, Ochiai et al, 1996;Ogasawara et al, 1998), DAP BII (Ogasawara, Kobayashi et al, 1996), DAP BIII (Ogasawara, Kobayashi et al, 1996) and DAP IV (Ogasawara, Ogawa et al, 1996;Ogasawara et al, 2005) from Pseudoxanthomonas mexicana WO24, which is a Gram-negative aerobic bacterium isolated from the wastewater of a bean curd (tofu) factory. On the basis of the enzymological data that we obtained, we proposed that bacterial DAPs should be classified in a manner different from that of mammalian DPPs, except for DAP IV (Ogasawara, Kobayashi et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

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Crystallization and preliminary X-ray crystallographic studies of dipeptidyl aminopeptidase BII from Pseudoxanthomonas mexicana WO24

et al. 2014

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Dipeptidyl aminopeptidase BII from Pseudoxanthomonas mexicana WO24 (DAP BII) is able to cleave a variety of dipeptides from the amino-terminus of substrate peptides. For crystallographic studies, DAP BII was overproduced in Escherichia coli, purified and crystallized using the hanging-drop vapourdiffusion method. X-ray diffraction data to 2.3 Å resolution were collected using an orthorhombic crystal form belonging to space group P2 1 2 1 2 1 , with unit-cell parameters a = 76.55, b = 130.86, c = 170.87 Å . Structural analysis by the multiwavelength anomalous diffraction method is in progress.

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Section: X-ray Analysessupporting

confidence: 82%

Section: Resultsmentioning

confidence: 99%

Section: Overproduction and Purificationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Crystallization and preliminary X-ray crystallographic studies of dipeptidyl aminopeptidase BII from Pseudoxanthomonas mexicana WO24

et al. 2014

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“…DAPs are present in all organisms, from bacteria to mammals, and the enzyme contributes to several important physiological functions such as the absorption of nutrients, immunity and the ability of pathogens to invade host cells151617. DAP BI18 and DAP IV19 show amino acid sequence homology and similar substrate specificity to enzymes that belong to the peptidase family S920 in clan SC, such as oligopeptidase B and DPP IV. In contrast, no homologs have been identified for DAP BII and DAP BIII.…”

mentioning

confidence: 99%

Identification of the Catalytic Triad of Family S46 Exopeptidases, Closely Related to Clan PA Endopeptidases

Suzuki

Sakamoto

Tanaka

et al. 2014

Sci Rep

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The exopeptidases of family S46 are exceptional, as the closest homologs of these enzymes are the endopeptidases of clan PA. The three-dimensional structure of S46 enzymes is unknown and only one of the catalytic residues, the serine, has been identified. The catalytic histidine and aspartate residues are not experimentally identified. Here we present phylogenetic and experimental data that identify all residues of the catalytic triad of S46 peptidase, dipeptidyl aminopeptidase BII (DAP BII) from Pseudoxanthomonas mexicana WO24. Phylogenetic comparison with the protein and S46 peptidases, revealed His-86, Ser-657, and five aspartate residues as possible catalytic residues. Mutation studies identified the catalytic triad of DAP BII as His-86, Asp-224, and Ser-657, while secondary structure analysis predicted an extended alpha-helical domain in between Asp-224 and Ser-657. This domain is unique for family S46 exopeptidases and its absence from the endopeptidases of clan PA might be key to their different hydrolysis activities.

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A luxR Homolog, aviR, in Agrobacterium vitis Is Associated with Induction of Necrosis on Grape and a Hypersensitive Response on Tobacco

Zheng¹,

Zhang²,

Carle³

et al. 2003

MPMI

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A Tn5 mutant of Agrobacterium vitis F2/5 (M1154) differs from the wild-type strain in that it has lost its abilities to cause necrosis on grape and a hypersensitive-like response (HR) on tobacco. The Tn5 insertion occurred in an open reading frame (ORF) aviR that is homologous to genes encoding the LuxR family of transcriptional regulators, thereby suggesting that the HR and necrosis are regulated by a quorum-sensing system. Fewer N-acyl-homoserine lactone autoinducers were detected in extracts from M1154 compared with extracts from F2/5 and from aviR-complemented M1154. The complemented mutant regained full ability to cause grape necrosis and HR. Eighteen ORFs located on a 36.6-kb insert in cosmid clone CPB221, which includes aviR, were sequenced and aligned with homologous genes from A. tumefaciens C58 and Sinorhizobium meliloti Rm1021. The order of several clustered genes is conserved among the bacteria; however, rearrangements are also apparent. Reverse transcriptase-polymerase chain reaction analysis indicated that ORF2 and ORF14 may be regulated by an aviR-encoded transcriptional regulator. Single site-directed mutations in each of the ORFs, however, had no effect on expression of HR or necrosis as compared with the wild-type parent.

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The gene encoding dipeptidyl aminopeptidase BI from Pseudomonas sp. WO24: cloning, sequencing and expression in Escherichia coli

Cited by 6 publications

References 24 publications

Crystallization and preliminary X-ray crystallographic studies of dipeptidyl aminopeptidase BII from Pseudoxanthomonas mexicana WO24

Crystallization and preliminary X-ray crystallographic studies of dipeptidyl aminopeptidase BII from Pseudoxanthomonas mexicana WO24

Identification of the Catalytic Triad of Family S46 Exopeptidases, Closely Related to Clan PA Endopeptidases

A luxR Homolog, aviR, in Agrobacterium vitis Is Associated with Induction of Necrosis on Grape and a Hypersensitive Response on Tobacco

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