A Novel Supersecondary Structure, Polyproline -turn Helices : Synthesis and Structure

一种新型超二级结构，聚脯氨酸转螺旋：合成和结构

• 批准号: 02044148
• 负责人: MATSUSHIMA Norio
• 金额: $ 3.07万
• 依托单位: School of Allied Health Professions, Sapporo Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02044148/
• 关键词: tandem repeats; computer graphics; structure prediction; nuclear magnetic resonance; small-angle x-ray scattering; supersecondary structure; 超二次構造; 縦列反復配列; コンピュ-タ-・グラフィックス; タコ・ロドプシン; グルテン; X線溶液散乱

During the past 5 years many protein sequences have been noted to contain unique tandem repeats having either a Tyr or Phe and 20-60% Pro with abundantly Gly, Gln, and Ser. Matsushima, Creutz and Kretsinger have developed a group of related helical models for seven proteins (molluscan rhodopsin, synaptophytsin, synexin, gliadin, RNA polymerase II, hordein, and gluten) based on the polyproline conformation combined with one or two beta-turns. In the present project we puropose to confirm the existence of polyproline, beta-turn helices using both theoretical and experimental methods.First method is the measurements of proton nuclear magnetic resonance (NMR) of synthetic repeat pentapeptide, (Tyr-Pro.Pro.Gln.Gly)n, of which the tandem repeat is contained at the carboxy-terminal domain of molluscan rhodopsin. Ac-(Tyr-Pro.Pro.Gln.Gly)n-NH_2 (N=2, 4, 6, and 8) was synthesized by solid-phase methods and was purified by HPLC. The two-dimensional NMR studies were carried out on DMS0-d_6 solutio … More ns of Ac-(Tyr.Pro.Pro.Gln.Gly)n-NH2 (n=2 and 4). Some inter-residue NOEs were observed, containing two NOEs between the Tyr alphaCH and the Pro deltaCH and the Pro deltaCH. Two observed NOEs indicate that bith the Tyr-Pro and the Pro-Pro bonds favor the trans-form in DMSO. The results of amide protons temperature coefficients, ^3J_Nalpha coupling constants, NOEs observed between inter-residues in Ac-(Tyr.Pro.Pro.Gln.Gly)_2-NH2 also indicated that Pro.Gln or Gly.Tyr may be involved in a type II beta-turn.Second method is small-angle x-ray scattering of one high molecular weight (HMW) subunit of wheat glutenin. The radius of gyration of whole particles, Ro, in aq. 50% (v/v) 1-propanol and 0/1M acetic acid 16.6 * 0.1nm and 22.8nm, respectively, and the corresponding radius of gyration of cross-section, Rc, was 2.82 * 0.02nm and 2.23 * 0.01nm, which indicate that the glutenin HMW subunit exists as very anisotropy particles in both solutions.Third method is molecular modeling by computer graphics and energy minimization for some tandem repeats. We tried to model the tandem repeats consisting of Pro.Gln.Gln.Pro.Phe.Pro.Gln within gamma-gliadin and Leu・Pro・Pro・Pro・Val・His within zein or glutelin-2 and developed a group of helical models closely related with polyproline beta-turn helices.These studies support that these tandem repeats take polyproline beta-turn helices, which are a fundamental basic supersecondary structure. Less

在过去的5年中，许多蛋白质序列被发现含有独特的串联重复序列，这些串联重复序列含有Tyr或Phe和20-60%的Pro，以及丰富的Gly，Gln和Ser。Matsushima，Creutz和Kretsinger已经为7种蛋白质开发了一组相关的螺旋模型（软体动物视紫红质，突触素，突触结合蛋白，麦醇溶蛋白，RNA聚合酶II，大麦醇溶蛋白，和麸质），其基于与一个或两个β-转角结合的聚脯氨酸构象。本课题的目的是从理论和实验两个方面来证实多聚脯氨酸β-转角螺旋的存在：第一种方法是对人工合成的重复五肽（Tyr-Pro.Pro.Gln.Gly）n进行质子核磁共振（NMR）测量，其中串联重复序列包含在软体动物视紫红质的羧基末端结构域。采用固相法合成了Ac-（Tyr-Pro.Pro.Gln.Gly）n-NH_2（N=2，4，6，8），经HPLC纯化。本文用二维核磁共振研究了DMS 0-d_6溶液的结构。 ...更多信息 Ac-（Tyr.Pro.Pro.Gln.Gly）n-NH2（n=2和4）的ns。观察到一些残基间NOE，在Tyr alphaCH和Pro deltaCH之间含有两个NOE，在Pro deltaCH之间含有两个NOE。观察到的两个NOE表明，在DMSO中，Tyr-Pro和Pro-Pro键有利于反式。Ac-（Tyr.Pro.Pro.Gln.Gly）_2-NH_2的酰胺质子温度系数、^3J_Nalpha偶合常数、氨基酸间NOE的测定结果也表明Pro.Gln或Gly.Tyr可能参与了II型β-转角。整个粒子的回转半径，Ro，在aq。50%（v/v）正丙醇和0/1 M乙酸的峰面积分别为16.6 × 0.1nm和22.8nm，相应的截面回转半径Rc分别为2.82 × 0.02nm和2.23 × 0.01nm，结果表明，谷蛋白HMW亚基在两种溶液中均以各向异性颗粒的形式存在一些串联重复的最小化。我们尝试模拟γ-麦醇溶蛋白中的Pro.Gln.Gln.Pro.Phe.Pro.Gln和玉米醇溶蛋白或谷蛋白-2中的Leu·Pro·Pro·Pro·瓦尔·His组成的串联重复序列，建立了一组与聚脯氨酸β转角螺旋密切相关的螺旋模型，这些研究支持这些串联重复序列具有聚脯氨酸β转角螺旋，这是一种基本的超二级结构。少

项目成果

MATSUSIMA,N.,Dinno,G.,SASAKI,N.,and IZUMI,Y: "Small-Augle X-Ray Scattering Study by Synchrotron Reveals That High Molecular Weight of Gutenin Is a Very Anisotropic Molecule" Biochem.Biophys.Res,Commun.186. 1057-1064 (1992)

MATSUSIMA,N.、Dinno,G.、SASAKI,N. 和 IZUMI,Y：“同步加速器的小口径 X 射线散射研究表明高分子量的古藤素是一种非常各向异性的分子”Biochem.Biophys.Res，

松嶋 範男: "新しい超二次構造をもつタンパク質" 高分子. 18. 524- (1990)

Norio Matsushima：“具有新的超二级结构的蛋白质”Polymer 18. 524-（1990）。

N. Matsushima, C. E. Creutz and R. H. Kretsinger: "Polyproline, -turn helices. Novel secondary structure proposed for the tandem repeats within rhodopsin, synaptophysin, synexin, gliadin, RNA polymerase II, hordein and gluten" Proteins : Structure, Functi

N. Matsushima、C. E. Creutz 和 R. H. Kretsinger：“聚脯氨酸，-转角螺旋。为视紫红质、突触素、突触素、麦醇溶蛋白、RNA 聚合酶 II、大麦醇溶蛋白和麸质内的串联重复提出的新型二级结构” 蛋白质：结构、功能

N. Matsushima, G. Danno, N. Sasaki, and Y. Izumi: "Small-angle x-ray scattering study by synchrotron orbital radiation reveals that high molecular weight of glutenin is a very anisotropic molecule" Biochem. Biophys. Res. Commun.186. 1057-1064 (1992)

N. Matsushima、G. Danno、N. Sasaki 和 Y. Izumi：“通过同步加速器轨道辐射进行的小角 X 射线散射研究表明，高分子量的麦谷蛋白是一种各向异性非常强的分子”Biochem。

松嶋 範男: "タンパク質の反復アミノ酸配列の立体構造予測:コンピュ-タ-・グラフィックスの利用" 生物物理. 31. 115-120 (1991)

Norio Matsushima：“蛋白质重复氨基酸序列的三维结构的预测：计算机图形学的使用”生物物理学。 31. 115-120 (1991)