Elucidation of Structural Effect of C-terminal Amidation of Bioactive Peptide

生物活性肽C端酰胺化结构效应的阐明

• 批准号: 14572041
• 负责人: IN Yasuko
• 金额: $ 1.92万
• 依托单位: Osaka University of Pharmaceutical Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14572041/
• 关键词: C-terminal amidation; endomorphin-2; dipeptides; solution structure; crystal structure; X-ray analysis; NMR; molecular dynamics simulation; C-terminal amidation; molecular conformation; X-ray analysis; crystal structure; solution conformation

1)X-ray crystal structure analyses of C-terminal amidated and nonamidated dipeptidesAs a series of elucidating the structural features of peptides caused by the C-terminal α-amidation, the crystal structures of H-Val-Gly-NH2, H-Ser-Phe-NH2, H-Gly-Tyr-NH2, and H-Pro-Tyr-NH2 hydrochloride salts were analyzed by X-ray diffraction method. Although respective molecules take the energetically allowable torsion angles concerning the backbone and side chains, their conformations are not necessarily the same as the corresponding unamidated ones. This is resulted from the different molecular packing requirement, rather than the different conformational feature inherent in the C-amidated and -unamidated peptides. As for the molecular packing feature, each peptide tends to form the repeated structure through the hydrogen bonds in which both amide NH and O=C groups participate. The chloride ions are located between the neighboring peptides and are hydrogen-bonded to the respective amide NHs, leadin … More g to the sheet structure. The hydrogen-bonding feature of amide group and its function for the molecular packing has been discussed based on the results so far analyzed.2)Analyses of solution structures of m-opioid agonist endomorphin 2 and its C-terminal OH formIn order to make clear the structural role of the C-terminal amide group of endomorphin-2 (EM2, H-Tyr-Pro-Phe-Phe-NH2), an endogenous m-receptor ligand, for the biological function, the solution conformations of endomorphin-2 and its C-terminal free acid (EM2OH, H-Tyr-Pro-Phe-Phe-OH), studied by two-dimensional ^1H-NMR measurements and molecular modeling calculations, were compared. Both peptides were in equilibrium between the cis and trans isomers around the Tyr-Pro w bond in a population of approximately 1:2 ratio in TFE and water and only trans in the membrane-mimetic micelles of perdeuterated dodecylphosphocholine (DPC). Fifty possible 3D structures for the each isomer were generated by the dynamical simulated annealing method under the proton-proton distance constraints derived from the ROE cross peaks. EM2 conformers adopt an open conformation in both TFE and water, whereas EM2OH shows conformational variation between the extended and folded backbone structures. On the other hand, EM2 in DPC takes the extended and folded backbone structures in 2:1 ratio, whereas EM2OH shows an open conformation. These results indicate clearly that the substitution of carboxyl group for C-terminal amide group makes the peptide flexible. The conformational requirement for μ-receptor activation has been discussed based on the active form proposed for endomorphin-1 and by comparing conformational features of EM2 and EM2OH. Less

1)C-端酰胺化和非酰胺化二肽的X-射线晶体结构分析作为对C-末端α-酰胺化产物结构特征的一系列阐明，用X射线衍射法分析了H-Val-Gly-NH_2、H-Ser-Phe-NH_2、H-Gly-Tyr-NH_2和H-Pro-Tyr-NH_2盐酸盐的晶体结构。虽然各自的分子相对于主链和侧链具有能量上允许的扭转角，但它们的构象不一定与相应的未酰胺化的构象相同。这是由于不同的分子包装要求，而不是C-酰胺化和未酰胺化多肽固有的不同构象特征所致。在分子堆积特征方面，每个肽都倾向于通过氨基NH和O=C参与的氢键形成重复结构。氯离子位于相邻的多肽之间，并与相应的酰胺NHS、先导…氢键连接对板材结构的要求更高。2)对m-阿片激动剂内吗啡-2及其C-端羟基形成的溶液结构进行了分析，以明确内源性m-受体配体--内吗啡-2的C-端酰胺基(EM2，H-Tyr-Pro-Phe-Phe-NH2)的结构作用。通过二维1H-核磁共振测量和分子模拟计算进行了比较。在TFE和水中，这两种多肽都处于Tyr-Pro w键附近的顺式和反式异构体之间，比例约为1：2，只有反式的十二烷基磷胆碱(DPC)在模拟膜的胶束中才是平衡的。用动力学模拟退火法在ROE交叉峰的质子-质子距离约束下生成了每个异构体的50个可能的三维结构。EM2构象在TFE和水中都采用开放构象，而EM2OH在伸展和折叠的主干结构之间表现出构象变化。另一方面，DPC中的EM2以2：1的比例呈伸展和折叠的主干结构，而EM2OH则呈开放构象。这些结果清楚地表明，羧基取代了C-端酰胺基，使多肽具有柔韧性。根据内吗啡素-1的活性形式，并通过比较μ和EM2OH的构象特征，讨论了内吗啡受体激活的构象要求。较少

项目成果

F.Yokokawa, H.Sameshima, Y.In, K.Minoura, T.Ishida, T.Shioiri: "Total synthesis and conformational studies of ceratospongamide, a bioactive cyclic heptapeptide from marine origin"Tetrahedron. 58. 8127-8143 (2002)

F.Yokokawa、H.Sameshima、Y.In、K.Minoura、T.Ishida、T.Shioiri：“来自海洋的生物活性环状七肽角海绵酰胺的全合成和构象研究”四面体。

Y.In, S.Kishima, K.Minoura, T.Nose, Y.Shimohigashi, T.Ishida: "Aggregation feature of fluorine-substituted benzene rings and intermolecular C-H...F interaction crystal structure analyses of mono-and trifluoro-L-phenylal anines"Chem.Pharm.Bull. 51(11). 125

Y.In、S.Kishima、K.Minoura、T.Nose、Y.Shimohigashi、T.Ishida：“氟取代苯环的聚集特征以及单氟和三氟的分子间 C-H...F 相互作用晶体结构分析

Y.In, H.Ohishi, T.Ishida, Y.Igarashi: "Concerted interaction between conjugated double bond CHs and multiple OHs in polyene macrolide antibiotic chainin : weak =C-H...O interactions responsible for intrinsic molecular assembly"Chem.Commun.. 14. 1692-1693

Y.In、H.Ohishi、T.Ishida、Y.Igarashi：“多烯大环内酯抗生素链中共轭双键 CH 和多个 OH 之间的协同相互作用：弱 = C-H...O 相互作用负责内在分子组装”Chem.Commun

Y.In, H.Ono, T.Ishida: "Structural studies on C-amidated -amino acids and peptides : function of amide group in molecular association in crystal structures of Val-Gly-NH2, Ser-Phe-NH2, Gly-Tyr-NH2 and Pro-Tyr-NH2 hydrochloride salts"Chem.Pharm.Bull.. 50.

Y.In、H.Ono、T.Ishida：“C-酰胺化氨基酸和肽的结构研究：酰胺基团在 Val-Gly-NH2、Ser-Phe-NH2、Gly- 晶体结构中分子缔合中的功能

A.Yamamoto, K.Tomoo, K.Mathugi, T.Hara, Y.In, T.Ishida: "Structural basis for development of cathepsin B-specific noncovalent-type inhibitor : crystalstructure of cathepsin B-E64c complex"Biochem.Biophys.Res.Commun.. 1597. 244-251 (2002)

A.Yamamoto、K.Tomoo、K.Mathugi、T.Hara、Y.In、T.Ishida：“开发组织蛋白酶 B 特异性非共价型抑制剂的结构基础：组织蛋白酶 B-E64c 复合物的晶体结构”Biochem.Biophys