MODULATION OF PROTEIN AND CELL FUNCTIONS BY HEPARIN/HEPARAN SULFATES

肝素/硫酸乙酰肝素对蛋白质和细胞功能的调节

• 批准号: 6108068
• 负责人: AUDREY L STONE
• 金额: --
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6108068
• 关键词: antiAIDS agent; anticoagulants; antiviral agents; chemical models; chemical structure function; heparan sulfate; heparin; human immunodeficiency virus 1; human tissue; nuclear magnetic resonance spectroscopy; virus infection mechanism

This project elucidates the fundamental molecular design of heparins and heparan sulfates (H/HS) and studies the structural basis of their specific modulations of protein and cell membrane in diverse normal and diseased physiological systems, using various biological, biochemical, and physical approaches. We examined H/HS as a modulator of viral infectivity, studied structure-function relations of a heparin-mimetic drug composed of a mixture of sulfated oligoxylans (S-oligoS) which, as does heparin, inhibits infectivity of HIV-1 in vitro: 1) We had shown that the capacity to inhibit HIV-1 cytotoxicity and syncytium-forming infectivity was governed by a structural specificity, and that inhibition of the two direct cell-killing mechanisms may have different structural requirements. A minimal-sized, potent antiviral component, CpF, was separable from S-oligoS having anticoagulation activity against thrombin and there was structural specificity in anticoagulation reactions. S-oligoS contained multiple D-G1cA moieties, suggesting a novel structural element, three beta 1, 4-linked xyloses with an beta 1,2-linked D-G1cA branch; 2) CpF-PkII isolated from CpF and further-purified CpF-PkII show that one or more sugars may be in the alternate chair form (axial as well as expected equatorial sulfates are found); 3) CpF inhibits the adherence of human CD4 cells to gp120-coated plastic, while S-oligoS that are inactive against HIV-1 in the above assays do not. CpF-PkI appears stable in size and charge during substitution and conjugation reactions. We are now able to develop a florescent CpF-PkI probe as a ligand for human CD4 cells to crosslink a putative S-oligoS "receptor" upon photoactivation; and 4) OligoS (8-, 12-, and 14-mers) of heparin purified by function (antithrombin activation) indicate that three turns about the axis is one requirement for capacity to form helical order, greater than or equal to 12- mer, as reelected by the geometry of bound cationic ligands.

该项目阐明了肝素和 乙par硫酸盐（H/HS）并研究其结构基础 蛋白质和细胞膜的特定调制在不同的正常和 患病的生理系统，使用各种生物学，生化， 和物理方法。 我们检查了H/HS作为病毒的调节剂 感染性，研究肝素模拟的结构 - 功能关系 由硫酸寡素（S-Oligos）的混合物组成的药物，该混合物是 肝素是否在体外抑制HIV-1的感染力：1）我们已经显示了 抑制HIV-1细胞毒性和合成形成的能力 感染性受结构特异性的控制，并受到抑制 在两个直接杀解细胞的机制中，可能具有不同的结构 要求。 最小尺寸的有效抗病毒成分CPF为 与具有抗凝活性的S-Oligos分离 并且在抗凝反应中存在结构特异性。 S-Oligos包含多个D-G1CA部分，暗示了一种新颖 结构元素，三个β1，带有β的4连接木质糖 1,2连接的D-G1CA分支； 2）从CPF分离的CPF-PKII和 进一步纯化的CPF-PKII表明一种或多种糖可能在 替代椅子形式（轴向和预期的赤道硫酸盐是 成立）; 3）CPF抑制了人CD4细胞对GP120涂层的依从性 塑料，而S-Oligos在上述中对HIV-1不活跃 测定没有。 CPF-PKI在大小和充电期间看起来稳定 替代和共轭反应。 我们现在能够开发 荧光CPF-PKI探针作为人CD4细胞交叉链接A的配体 假定的S-Oligos“受体”光活化；和4）寡素（8-，， 通过功能纯化的肝素的12-和14-mers）（抗凝血酶 激活）表明轴是三个转弯是一个要求 为了形成大于或等于12 mer的螺旋顺序的能力 通过结合阳离子配体的几何形状转率。