Three-dimensional structure and Molecular Evolution of Tandem repeals within Ice Nucleation Proteins

冰成核蛋白内串联废除的三维结构和分子进化

• 批准号: 13680746
• 负责人: MATSUSHIMA Norio
• 金额: $ 1.66万
• 依托单位: Sapporo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13680746/
• 关键词: Ice nucleation protein; Tandem repeats; Loop structure; NMR; Glycine-rich proteins; Molecular evolution; 氷核タンパク質; タンデムリピート; 核磁気共鳴法(NMR)

Certain bacteria nucleate the crystallization of ice. The ice-nucleation activity is conferred by a protein (ice nucleation protein ; INP). The amino acid sequences of the INPs with molecular weights near 120kDa have a central repeating domain (comprising approximately 81% of the total sequence). The repeating domain consists of tandem repeats with the consensus sequence of AxxxSxxx. The structure of the INPs is still unknown. The purpose of the present study is to investigate the evolution and the structure of the INPs. First, we performed, comparative sequence analysis of ten INPs in details. The analysis revealed that the repeating domain is separated into four subdomains (R^c, R^1, R^2 and R^N). On the basis of this result, the evolutionary history was discussed. Secondly, we performed NMR experiments of synthetic peptide H-SGLRSVLTAGYGSSLISGRRSSLT-OH corresponding to sections of the R^N subdomain. The NMR results indicated the presence of a loop conformation in the sequence of LTAGY.

某些细菌使冰结晶成核。冰成核活性由蛋白质（冰成核蛋白；INP）赋予。分子量接近120kDa的INP的氨基酸序列具有中心重复结构域（约占总序列的81%）。重复结构域由具有 AxxxSxxx 共有序列的串联重复序列组成。 INP 的结构仍然未知。本研究的目的是研究 INP 的进化和结构。首先，我们对十个INP进行了详细的比较序列分析。分析表明，重复结构域分为四个子结构域（R^c、R^1、R^2 和 R^N）。在此结果的基础上，讨论了进化历史。其次，我们对对应于 R^N 子结构域部分的合成肽 H-SGLRSVLTAGYGSSLISGRRSSLT-OH 进行了 NMR 实验。 NMR结果表明LTAGY序列中存在环构象。

项目成果

Hayashi N, Matsubara M, Jinbo Y, Titani K, Izumi Y, Matsushima N.: "Nef of HIV-1 interacts directly with calcium-bound calmodulin"Protein Sci. 11(3). 529-537 (2002)

Hayashi N、Matsubara M、Jinbo Y、Titani K、Izumi Y、Matsushima N.：“HIV-1 的 Nef 直接与钙结合钙调蛋白相互作用”Protein Sci。

Kumaki Y, Matsushima N, Yoshida H, Nitta K, Hikichi K.: "Structure of the YSPTSPS repeat containing two SPXX motifs in the CTD of RNA polymerase II. NMR studies of cyclic model peptides reveal that SPTS turn is more stable than SPSY in water"Biochim. Biop

Kumaki Y、Matsushima N、Yoshida H、Nitta K、Hikichi K.：“RNA 聚合酶 II 的 CTD 中包含两个 SPXX 基序的 YSPTSPS 重复结构。环状模型肽的 NMR 研究表明，SPTS 转角比 SPSY 更稳定