Design of tailor-made micro-carrier protein having molecular-selective recognition.

具有分子选择性识别的定制微载体蛋白的设计。

• 批准号: 17300165
• 负责人: INUI Takashi
• 金额: $ 9.54万
• 依托单位: Osaka Prefecture University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17300165/
• 关键词: transporter protein; drug delivery system; lipocalin family protein; molecular-selective recognition; hydrophobic ligand; ドラッグデリバリーシステム

Small-angle X-ray scattering (SAXS) of lipocalin-type prostaglandin D synthase (L-PGDS) and two other lipocalins, β-lactoglobulin (βLG) and retinal-binding protein (RBP) were measured to clarify their conformational changes induced by binding of small lipophilic ligands, such as all-trans-retinoic acid (RA), bilirubin (BR) and biliverdin (BV). The radius of gyration (R_g) was estimated to be 19.4Å for L-PGDS, and 18.8Å for L-PGDS/RA, 17.3Å for L-PGDS/BR and 17.8Å for L-PGDSn3V complexes, indicating that L-PGDS became compact after binding of these ligands. Alternatively, the R_g of βLG and RBP (20.3 and 26.2Å, respectively) was almost unchanged before and after binding of RA. Molecular modeling of L-PGDS based on the SAXS data indicated that the ligand binding changed the peripheral region of the molecule and suggested that such structural flexibility of the L-PGDS molecule is responsible for the broader ligand selectivity of L-PGDS than other lipocalins.The three-dimensional structure of recombinant mouse L-PGDS was determined by NMR. The structure of L-PGDS exhibited the typical lipocalin-fold, consisting of an 8-stranded, antiparallel β-barrel and a α-helix associated with the outer surface of the barrel. The interior of the barrel formed a hydrophobic cavity opening to the upper end of the barrel, the size of which was larger than those of other lipocalins and the cavity contained two pockets. Molecular docking studies based on the result of NMR experiments with RA and PGH_2 analog revealed that PGH_2 fully occupied the hydrophilic pocket 1, where Cys65 was located, and RA did the hydrophobic pocket 2, where amino acid residues important for retinoid-binding in other lipocalins were well conserved. These results indicated that the two binding sites in the large cavity of L-PGDS was responsible for the broad ligand specificity of L-PGDS and the non-competitive inhibition of L-PGDS activity by RA.

用小角X射线散射（SAXS）方法研究了脂笼蛋白型前列腺素D合成酶（L-PGDS）、β-乳球蛋白（β-lactoglobulin，βLG）和视黄结合蛋白（retinal-binding protein，RBP）与全反式维甲酸（all-trans-retinoic acid，RA）、胆红素（bilirubin，BR）和胆绿素（biliverdin，BV）等亲脂配体结合后的构象变化。L-PGDS/RA、L-PGDS/BR和L-PGDSn 3V复合物的回转半径分别为19.4、18.8、17.3和17.8 μ m，表明L-PGDS与这些配体结合后变得紧密。β-LG和RBP的R_g在与RA结合前后几乎没有变化（分别为20.3和26.2 μ g）。基于SAXS数据的L-PGDS的分子模拟表明，配体结合改变了分子的外围区域，并表明L-PGDS分子的这种结构灵活性是L-PGDS比其他脂质运载蛋白更广泛的配体选择性的原因。L-PGDS的结构表现出典型的脂质运载蛋白折叠，由一个8链的反平行β-桶和一个与桶外表面相关的α-螺旋组成。筒的内部形成了一个疏水性空腔，开口朝向筒的上端，其尺寸大于其他脂质运载蛋白的尺寸，并且空腔包含两个口袋。基于核磁共振实验结果的分子对接研究表明，PGH_2完全占据了亲水口袋1中Cys 65的位置，而RA占据了疏水口袋2中其他脂质运载蛋白中与类维生素A结合的重要氨基酸残基。这些结果表明，L-PGDS大空腔中的两个结合位点是L-PGDS广泛的配体特异性和RA对L-PGDS活性的非竞争性抑制的原因。

项目成果

Thermal stability of lipocalin-type prostaglandin D synthase induced by hydrophobic small ligand.

疏水性小配体诱导的脂质运载蛋白型前列腺素 D 合酶的热稳定性。

• 发表时间: 2005
• 期刊: The society of the study for life and environmental science at Tsu City College 53
• 作者: Saitoh;R.;Suzuki;T.;Yoneyama;T.;Mutoh;S.;Hasegawa;K.;Munehiro;M.;Niwa;K.;Watanabe;T.;Ohyama;T;Toshihiro Watanabe;Daisuke Irikura;Yoshitaka Nagai;Daisuke Irikura;Yoshitaka Nagai;Ikuko Mohri;Ko Fujimori;Osamu Ishibashi;飯田津喜美;Ikuko Mohri;Tsukimi Iida;Ikuko Mohri;飯田津喜美;Ko Fujimori;Tsukimi Iida;Bruno Kilunga Kubata;西口茉実;Bruno Kilunga Kubata;Tsukimi Iida;Nishiguchi Mami
• 通讯作者: Nishiguchi Mami

疎水性低分子結合によるリポカリン型プロスタグランジンD合成酵素の熱安定性についての研究

疏水性小分子结合研究脂质运载蛋白型前列腺素D合酶的热稳定性

• 发表时间: 2005
• 期刊: 三重短期大学生活科学研究会(紀要) 第53号
• 作者: Saitoh;R.;Suzuki;T.;Yoneyama;T.;Mutoh;S.;Hasegawa;K.;Munehiro;M.;Niwa;K.;Watanabe;T.;Ohyama;T;Toshihiro Watanabe;Daisuke Irikura;Yoshitaka Nagai;Daisuke Irikura;Yoshitaka Nagai;Ikuko Mohri;Ko Fujimori;Osamu Ishibashi;飯田津喜美;Ikuko Mohri;Tsukimi Iida;Ikuko Mohri;飯田津喜美;Ko Fujimori;Tsukimi Iida;Bruno Kilunga Kubata;西口茉実
• 通讯作者: 西口茉実

Characterization of a major secretory protein in the cane toad (Bufo marinus) choroid plexus as an amphibian lipocalin-type prostaglandin D synthase.

甘蔗蟾蜍 (Bufo marinus) 脉络丛中主要分泌蛋白的表征，为两栖类脂质运载蛋白型前列腺素 D 合酶。

• 发表时间: 2007
• 期刊: J. Biochem. 141
• 作者: Saitoh;R.;Suzuki;T.;Yoneyama;T.;Mutoh;S.;Hasegawa;K.;Munehiro;M.;Niwa;K.;Watanabe;T.;Ohyama;T;Toshihiro Watanabe;Daisuke Irikura
• 通讯作者: Daisuke Irikura

Structural and mutational analysis of Trypanosoma brucei prostaglandin H_2 reductase provides insight into the catalytic mechanism of aldo-ketoreductases.

布氏锥虫前列腺素 H_2 还原酶的结构和突变分析有助于深入了解醛酮还原酶的催化机制。

• 发表时间: 2005
• 期刊: J.Biol.Chem. 280
• 作者: Saitoh;R.;Suzuki;T.;Yoneyama;T.;Mutoh;S.;Hasegawa;K.;Munehiro;M.;Niwa;K.;Watanabe;T.;Ohyama;T;Toshihiro Watanabe;Daisuke Irikura;Yoshitaka Nagai;Daisuke Irikura;Yoshitaka Nagai;Ikuko Mohri;Ko Fujimori;Osamu Ishibashi;飯田津喜美;Ikuko Mohri;Tsukimi Iida;Ikuko Mohri;飯田津喜美;Ko Fujimori;Tsukimi Iida;Bruno Kilunga Kubata;西口茉実;Bruno Kilunga Kubata
• 通讯作者: Bruno Kilunga Kubata

Thermal unfolding mechanism of lipocalin-type prostaglandin D synthase induced by hydrophobic small ligand.

疏水性小配体诱导脂质运载蛋白型前列腺素 D 合酶的热解折叠机制。

• 发表时间: 2006
• 期刊: The society of the study for life and environmental science at Tsu City College 54
• 作者: Saitoh;R.;Suzuki;T.;Yoneyama;T.;Mutoh;S.;Hasegawa;K.;Munehiro;M.;Niwa;K.;Watanabe;T.;Ohyama;T;Toshihiro Watanabe;Daisuke Irikura;Yoshitaka Nagai;Daisuke Irikura;Yoshitaka Nagai;Ikuko Mohri;Ko Fujimori;Osamu Ishibashi;飯田津喜美;Ikuko Mohri;Tsukimi Iida
• 通讯作者: Tsukimi Iida