Novel Peptides Against Modified Heparan Sulfate

抗修饰硫酸乙酰肝素的新肽

• 批准号: 8702500
• 负责人: DEEPAK SHUKLA
• 金额: $ 24.66万
• 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 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8702500
• 关键词: Affinity; Anabolism; Antiviral Agents; Bacteriophages; Binding; Binding Sites; Biological Process; Cell fusion; Cell surface; Cells; Chemicals; Commit; Communicable Diseases; Complex; Dendrimers; Detection; Development; Family; Future; Generations; Glycobiology; Glycoproteins; 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; public health relevance; 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）的附着以及膜融合受体的能力与其结构和功能多样性有关，所述多样性源自其生物合成期间的广泛修饰。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。已经鉴定了3-OST家族的七个成员，并且已经表明3-OST中的每一个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各自独特的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 mer噬菌体展示文库筛选未修饰的HS和由3-O-磺基转移酶-3（3-OST-3）产生的3-OS HS，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对分离的G1和G2肽进行了表征。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，在初始筛选期间鉴定的几种抗HS和抗30 HS肽尚未针对HSV-1感染进行测试。我们的假设是，抗HS和抗3-OS HS肽的整个面板的表征将不仅增加我们的机会，以分离最有效的抑制剂，分别对HS和修饰的HS，但它也将导致开发新的试剂，以映射出结构复杂的HS链。本研究的目的1是合成抗HSV-1侵入和细胞间扩散的3-O-硫酸乙酰肝素（3-OS HS）结合肽，并对其功能进行表征。我们将生成关于单个肽干扰HSV-1附着、进入和细胞间传播的能力的详细信息。目的2将确定3- OS HS肽针对多种3-OST同种型的抗HSV潜力。它还将确定肽对3种抗体的相对交叉特异性。 OS HS由其他同种型产生。我们的研究将产生一些有价值的试剂，针对各种3-OS HS修改，并大大有助于了解HS在人类生物学和感染性疾病中的功能。