NMR studies of human interleukin-6 and its mutants

人白细胞介素 6 及其突变体的 NMR 研究

• 批准号: 07672310
• 负责人: NISHIMURA Chiaki
• 金额: $ 1.41万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07672310/
• 关键词: IL-6; NMR; DSC; CD; Mutants; Structure-function relationship; Structive-stability relationship

On the basis of the partial signal assignments and the observed NOE network, the folding topology of human IL-6 was analyzed. A comparison of the folding topology of IL-6 with that of human G-CSF indicated that IL-6 has a significant similarity of folding topology to that of G-CSF.DSC thermogram of the wild-type IL-6 at pH 4.2 showed two endothermic peaks at 35 and 65ﾟC,indicating that an intermediate state exists in the heat-denaturation pathway. In order to understand the structure-function and structure-stability relationships in the human IL-6 system, comparative studies were performed on the basis of NMR,DSC,and CD data obtained using the wild-type IL-6 and six mutants (L152V,L159V,L166V,L168V,L175V,and L182V). The NMR data showed that L182V substitution induced no structural change in IL-6, suggesting that Leu182 is located on the surface of the IL-6 molecule. A significant decrease in receptor-binding activity was observed in the L182V mutant. It was concluded that the side-chai … More n of Leu182 is directly involved in receptor-binding. L175V substitution was shown to induce a significant structural change in IL-6. It is possible that helix D bent more sharply toward helix B in the L175V mutant than in the wild-type IL-6 to maintain a closely packed and solvent-inaccessible core formed in the mutated region. It is suggested that the kink of helix D is related to the decrease in receptor-binding activity in the L175V mutant. In the case of L152V mutant, a significant structural changes were observed compared with the wild-type IL-6. However, no difference in receptor-binding activity between the wild-type IL-6 and L152V mutant was observed. The DSC data revealed that the partial unfolding of L152V mutant and the full unfolding of L175V mutant occurred at temperatures lower than those of the wild-type IL-6. The NMR data observed at various temperatures showed that L152V and L175V mutants are prone to the soluble self-association and insoluble precipitation, respectively, compared with the wild-type IL-6. Less

在部分信号归属和观察到的NOE网络的基础上，分析了人IL-6的折叠拓扑结构。比较IL-6和人G-CSF的折叠拓扑结构，发现IL-6和G-CSF的折叠拓扑结构有很大的相似性，野生型IL-6在pH4.2时的DSC热分析显示在35 ℃和65 ℃有两个吸热峰，表明在热变性过程中存在一个中间状态。为了理解人IL-6系统中的结构-功能和结构-稳定性关系，基于使用野生型IL-6和六种突变体（L152 V、L159 V、L166 V、L168 V、L175 V和L182 V）获得的NMR、DSC和CD数据进行比较研究。NMR数据显示，L182 V取代没有引起IL-6的结构变化，表明Leu 182位于IL-6分子的表面。在L182 V突变体中观察到受体结合活性显著降低。得出结论，侧链 ...更多信息 Leu 182的n直接参与受体结合。L175 V取代显示诱导IL-6的显著结构变化。可能的是，在L175 V突变体中，螺旋D比在野生型IL-6中更急剧地向螺旋B弯曲，以维持在突变区域中形成的紧密堆积和溶剂不可及的核心。这表明，螺旋D的扭结与L175 V突变体中受体结合活性的降低有关。在L152 V突变体的情况下，与野生型IL-6相比观察到显著的结构变化。然而，没有观察到野生型IL-6和L152 V突变体之间的受体结合活性的差异。DSC数据显示，L152 V突变体的部分解折叠和L175 V突变体的完全解折叠发生在低于野生型IL-6的温度下。在不同温度下观察到的NMR数据表明，与野生型IL-6相比，L152 V和L175 V突变体分别倾向于可溶性自缔合和不溶性沉淀。少

项目成果

西村千秋: "IL-6の高次構造" 臨床免疫. 27. 990-996 (1995)

Chiaki Nishimura：“IL-6 的高级结构”临床免疫学 27. 990-996 (1995)。

I.C.Nishimura, A.Watanabe, H.Gouda, I.Shimada, and Y.Arata: "Folding Topologies of Human Interleukin-6 and Its Mutants As Studied by NMR Spectroscopy" Biochemistry. 35. 273-281 (1996)

I.C.Nishimura、A.Watanabe、H.Gouda、I.Shimada 和 Y.Arata：“通过核磁共振波谱研究的人白细胞介素 6 及其突变体的折叠拓扑”生物化学。

Chiaki・Nishimura: "Folding Topologies of Human Interleukin-6 and Its Mutants As Studied by NMR Spectroscopy" Biochemistry. 35. 273-281 (1996)

Chiaki·Nishimura：“通过 NMR 光谱研究人类 Interleukin-6 及其突变体的折叠拓扑”，生物化学 35. 273-281 (1996)。

Nishimura,C.: "Folding Topologies of Human Interleukin-6 and Its Mutants As Studied by NMR Spectroscopy" Biochemistry. 35. 273-281 (1996)

Nishimura,C.：“通过核磁共振波谱研究人类白细胞介素 6 及其突变体的折叠拓扑”生物化学。