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Periplasmic form of dipeptidyl aminopeptidase IV from Pseudoxanthomonas mexicana WO24: purification, kinetic characterization, crystallization and X‐ray crystallographic analysis
Author(s) -
Roppongi Saori,
Tateoka Chika,
Fujimoto Mayu,
Iizuka Ippei,
Morisawa Saori,
Nakamura Akihiro,
Honma Nobuyuki,
Suzuki Yoshiyuki,
Shida Yosuke,
Ogasawara Wataru,
Tanaka Nobutada,
Sakamoto Yasumitsu,
aka Takamasa
Publication year - 2017
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x17014911
Subject(s) - periplasmic space , dipeptidyl peptidase , chemistry , aminopeptidase , tripeptide , crystallization , crystallography , escherichia coli , enzyme , biochemistry , peptide , amino acid , leucine , organic chemistry , gene
Dipeptidyl aminopeptidase IV (DAP IV or DPP IV) from Pseudoxanthomonas mexicana WO24 (PmDAP IV) preferentially cleaves substrate peptides with Pro or Ala at the P1 position [NH 2 ‐P2‐P1(Pro/Ala)‐P1′‐P2′…]. For crystallographic studies, the periplasmic form of PmDAP IV was overproduced in Escherichia coli , purified and crystallized in complex with the tripeptide Lys‐Pro‐Tyr using the hanging‐drop vapour‐diffusion method. Kinetic parameters of the purified enzyme against a synthetic substrate were also determined. X‐ray diffraction data to 1.90 Å resolution were collected from a triclinic crystal form belonging to space group P 1, with unit‐cell parameters a = 88.66, b = 104.49, c = 112.84 Å, α = 67.42, β = 68.83, γ = 65.46°. Initial phases were determined by the molecular‐replacement method using Stenotrophomonas maltophilia DPP IV (PDB entry 2ecf) as a template and refinement of the structure is in progress.