Novel Peptides Against Modified Heparan Sulfate

抗修饰硫酸乙酰肝素的新肽

• 批准号: 8917086
• 负责人: DEEPAK SHUKLA
• 金额: $ 19.3万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8917086
• 关键词: Affinity; Anabolism; Antiviral Agents; Bacteriophages; Binding; Binding Sites; Biological Process; Cell fusion; Cell surface; Cells; Chemicals; Communicable Diseases; Complex; Dendrimers; Detection; Development; Family; Future; Generations; Glycobiology; Glycoproteins; Health; Heparan Sulfate Biosynthesis; Heparitin Sulfate; Herpesvirus 1; Herpetic Keratitis; Human Biology; In Vitro; Individual; Infection; Inflammatory; Inorganic Sulfates; Lead; Letters; Libraries; Maps; Mediating; Membrane Fusion; Membrane Proteins; Modification; Molecular; Noise; Peptides; Phage Display; Polysaccharides; Protein Isoforms; Proteins; Reagent; Relative (related person); Signal Transduction; Simplexvirus; Site; Source; Specificity; Structure; Testing; Toxic effect; Unspecified or Sulfate Ion Sulfates; Virus Diseases; antimicrobial; cytokine; cytotoxicity; fighting; genital herpes; in vivo; in vivo Model; inhibitor/antagonist; macromolecule; member; microbial; microorganism interaction; novel; pathogen; practical application; receptor; screening; sulfation; sulfotransferase; tool

DESCRIPTION (provided by applicant): Cell surface heparan sulfate (HS) interacts with diverse proteins including the microbial surface proteins involved in attachment and/or entry into the host. The ability of HS to act as an attachment as well as membrane fusion receptor for herpes simplex virus (HSV) relates to its structural and functional diversity originating from extensive modifications during its biosynthesis. The final modification of HS is the 3-O sulfation which is mediated by 3-O-sulfotransferases (3-OSTs). Seven members of the 3-OST family have been identified and it has been suggested that each of the 3-OSTs are capable of recognizing unique saccharide sequences around the modification sites. This site-specific function of each isoform allows them to generate their own distinct 3-O-sulfated heparan sulfate (3-OS HS) motifs. In order to identify small peptide inhibitors of HS and 3-OS HS functions especially during HSV-1 entry, we screened both unmodified HS and 3-OS HS generated by 3-O-sulfotransferase-3 (3-OST-3) using 12 mer-phage display library and isolated a panel of peptides with unique sequence diversity. Of which, we characterized the G1 and G2 peptides isolated against unmodified HS and modified HS respectively. The characterization of G1 and G2 peptides demonstrated promising results by inhibiting HSV-1 infections both in vitro and in vivo models of ocular HSV-1 infection and genital herpes infection. However, several of the anti-HS and anti-3O HS peptides identified during initial screening have not been tested against HSV-1 infection. Our hypothesis is that characterization of the entire panel of anti-HS and anti-3-OS HS peptide will not only increase our chance to isolate the most potent inhibitor against HS and modified HS respectively but it will also lead towards the development of novel reagents to map out structurally complex chains of HS. Aim 1 of the proposal will focus on synthesis and functional characterization of anti-3-O-sulfated heparan sulfate (3-OS HS) binding peptides against HSV-1 entry and cell-to-cell spread. We will generate detailed information on the abilities of the individual peptides to interfere with HSV-1 attachment, entry and cell-to-cell spread. Aim 2 will determine the anti-HSV potential of 3- OS HS peptides against multiple 3-OST isoforms. It will also determine the relative cross specificities of the peptides against the 3 OS HS generated by other isoforms. Our study will produce some valuable reagents against various 3-OS HS modifications and help greatly with the understanding of HS functions in human biology and infectious diseases.

描述（由申请人提供）：细胞表面硫酸乙酰肝素（HS）与多种蛋白质相互作用，包括参与附着和/或进入宿主的微生物表面蛋白质。 HS 作为单纯疱疹病毒 (HSV) 附着和膜融合受体的能力与其生物合成过程中广泛修饰所产生的结构和功能多样性有关。 HS 的最终修饰是由 3-O-磺基转移酶 (3-OST) 介导的 3-O 硫酸化。 3-OST 家族的七个成员已被鉴定，并且表明每个 3-OST 都能够识别修饰位点周围的独特糖序列。每种亚型的这种位点特异性功能使它们能够产生自己独特的 3-O-硫酸化硫酸乙酰肝素 (3-OS HS) 基序。为了鉴定 HS 和 3-OS HS 功能的小肽抑制剂，特别是在 HSV-1 进入期间，我们使用 12 筛选了未修饰的 HS 和 3-O-sulfotransferase-3 (3-OST-3) 产生的 3-OS HS。 mer-phage 展示文库并分离出一组具有独特序列多样性的肽。其中，我们分别针对未修饰的 HS 和修饰的 HS 分离了 G1 和 G2 肽。 G1 和 G2 肽的表征通过在眼部 HSV-1 感染和生殖器疱疹感染的体外和体内模型中抑制 HSV-1 感染证明了有希望的结果。然而，在初步筛选过程中鉴定出的几种抗 HS 和抗 3O HS 肽尚未针对 HSV-1 感染进行测试。我们的假设是，整个抗 ​​HS 和抗 3-OS HS 肽组的表征不仅会增加我们分别分离最有效的 HS 和修饰 HS 抑制剂的机会，而且还将导致新型试剂的开发绘制结构复杂的 HS 链。该提案的目标 1 将重点关注针对 HSV-1 进入和细胞间传播的抗 3-O-硫酸化硫酸乙酰肝素 (3-OS HS) 结合肽的合成和功能表征。我们将生成有关各个肽干扰 HSV-1 附着、进入和细胞间传播的能力的详细信息。目标 2 将确定 3-OS HS 肽针对多种 3-OST 亚型的抗 HSV 潜力。它还将确定肽针对 3 种肽的相对交叉特异性。 OS HS 由其他亚型生成。我们的研究将产生一些针对各种 3-OS HS 修饰的有价值的试剂，并极大地帮助了解 HS 在人类生物学和传染病中的功能。

项目成果

Liposome-Mediated Herpes Simplex Virus Uptake Is Glycoprotein-D Receptor-Independent but Requires Heparan Sulfate.

脂质体介导的单纯疱疹病毒摄取不依赖于糖蛋白-D 受体，但需要硫酸乙酰肝素。

• 发表时间: 2016
• 作者: Burnham, Lorrie A;Jaishankar, Dinesh;Thompson, Jeffrey M;Jones, Kevin S;Shukla, Deepak;Tiwari, Vaibhav
• 通讯作者: Tiwari, Vaibhav