Roppongi Saori, Tateoka Chika, Fujimoto Mayu, Iizuka Ippei, Morisawa Saori, Nakamura Akihiro, Honma Nobuyuki, Suzuki Yoshiyuki, Shida Yosuke, Ogasawara Wataru, Tanaka Nobutada, Sakamoto Yasumitsu, Nonaka Takamasa
School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuta, Yahaba, Iwate 028-3694, Japan.
Department of Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata 940-2188, Japan.
Acta Crystallogr F Struct Biol Commun. 2017 Nov 1;73(Pt 11):601-606. doi: 10.1107/S2053230X17014911. Epub 2017 Oct 23.
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-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 P1, 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.
来自墨西哥假黄单胞菌WO24的二肽基肽酶IV（DAP IV或DPP IV，即PmDAP IV）优先切割在P1位置带有Pro或Ala的底物肽[NH-P2-P1(Pro/Ala)-P1'-P2'…]。为了进行晶体学研究，PmDAP IV的周质形式在大肠杆菌中过量表达，经纯化后，使用悬滴气相扩散法与三肽Lys-Pro-Tyr形成复合物并结晶。还测定了纯化酶对合成底物的动力学参数。从属于空间群P1的三斜晶体形式收集到了分辨率为1.90 Å的X射线衍射数据，晶胞参数为a = 88.66、b = 104.49、c = 112.84 Å，α = 67.42、β = 68.83、γ = 65.46°。初始相位通过分子置换法确定，使用嗜麦芽窄食单胞菌DPP IV（PDB条目2ecf）作为模板，并正在对结构进行优化。
