Investigating Complex Glycans on Biological Surfaces

研究生物表面上的复杂聚糖

• 批准号: 8072127
• 负责人: RAM SASISEKHARAN
• 金额: $ 19.4万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-19 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8072127
• 关键词: Address; Binding; Biochemical; Biological; Biological Process; Biology; Cell Communication; Cell Line; Cell surface; Cells; Cellular Tropism; Chemicals; Complex; Engineering; Environment; Epithelial; Event; Glycoside Hydrolases; Grant; Growth and Development function; Hemagglutinin; Human; Infection; Influenza A virus; Lectin; Link; Lung; Maps; Mass Spectrum Analysis; Mediating; Membrane Fusion; Membrane Glycoproteins; Modification; Molecular; Pathogenesis; Pattern; Physiological; Plant Lectins; Play; Polysaccharides; Property; Proteins; Role; Sialic Acids; Site; Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Staining method; Stains; Structure; Structure of respiratory epithelium; Structure-Activity Relationship; Surface; System; Testing; Tissue Stains; Tissues; Upper respiratory tract; Viral; Viral Pathogenesis; Virus Diseases; abstracting; base; cell type; extracellular; flexibility; immunoregulation; microbial; pathogen; public health relevance; receptor; receptor binding; respiratory; therapeutic development; tissue tropism; tool

DESCRIPTION (provided by applicant): Complex glycans, which are ubiquitously distributed at the interface of intercellular surfaces, act as receptors for various microbial pathogens and play a key role in determining the pathogen's hostcellular and tissue tropism. Despite their importance in host-pathogen interaction, it has been challenging to bridge the biochemical and biophysical specificity of glycan-protein interactions with the biological functions modulated by these interactions. These challenges arise from the difficulties in characterizing the fine structural granularity of glycan receptors in their physiological context such as epithelial surface of different tissues. Therefore there is a need to bridge the structural characterization of glycans at the tissue, cellular and molecular levels. Decoding the diversity of the physiological glycan receptors at these levels would not only facilitate the identification of glycan receptor targets from the standpoint of understanding pathogenesis but would also facilitate development of therapeutic strategies to counter pathogen infection. We have begun to address the above issues by developing a multifaceted framework to decode the structure-function relationship of glycan-protein interactions. Using a combination of lectin staining of tissues and mass spectrometry based structural profiling of glycans isolated from representative cell types we have taken a step towards bridging the characterization of glycans at the tissue and cellular level. Plant lectins have traditionally been used to investigate distribution of glycan structural motifs (recognized by the lectins) in the physiological context of cell and tissue surfaces. However, the diversity of glycan receptors based on lectin staining have been characterized in a abstract fashion or at best in terms of terminal glycan linkages such as a2-3 or a2-6 linked sialic acids. The abundant structural and biochemical information on glycan-binding properties of plant lectins offers tremendous potential to develop new strategies for using these lectins to probe into finer structural details of physiological glycans going beyond terminal linkages. Based on this information, lectins can be engineered to target interactions between pathogens and their host glycan receptors. Building on our previous efforts in this proposal we seek to develop strategies that utilize plant lectins in new ways to obtain more detailed structural characterization of physiological glycan receptors as well as develop these lectins to target pathogen-host glycan receptor interactions. PUBLIC HEALTH RELEVANCE: Complex glycans decorate biological surfaces specifically the interface of intercellular surfaces where they play a critical role in mediating cell - cell interactions such as host cell - pathogen interactions. There are challenges in bridging the physiological glycan receptor binding of pathogens with the biology of their pathogenesis and host adaptation. These challenges arise from the limitations in characterizing the fine structural diversity of glycan receptors in the appropriate physiological context such as specific cell and tissue surfaces. In this proposal we seek to address this issue by exploiting the untapped potential of plant lectins both to obtain a detailed mapping of the nuances in the structural features of the glycan receptors in physiological surfaces such as human upper respiratory epithelia and to develop these lectins as tools to competitively target these receptors.

描述（由申请人提供）：复杂的聚糖，它们在细胞间表面的界面上分布在多种微生物病原体的受体中，并在确定病原体的宿主细胞和组织疗法方面起关键作用。尽管它们在宿主 - 病原体相互作用中的重要性，但桥接聚糖 - 蛋白质相互作用与这些相互作用调节的生物学功能的生化和生物物理特异性的挑战。这些挑战是由于在生理环境中（例如不同组织的上皮表面）表征聚糖受体的精细结构颗粒性的困难引起的。因此，需要在组织，细胞和分子水平上弥合聚糖的结构表征。从理解发病机理的角度来看，在这些水平上解码生理聚糖受体的多样性不仅会促进聚糖受体靶标的鉴定，而且还将促进与病原体感染相反的治疗策略的发展。我们已经开始通过开发一个多方面的框架来解决上述问题，以解码聚糖 - 蛋白质相互作用的结构功能关系。使用从代表性细胞类型分离的组织的组织的凝集素染色和基于质谱的基于质谱的结构分析，我们朝着弥合组织和细胞水平的聚糖表征迈出了一步。 传统上，植物凝集素用于研究细胞和组织表面的生理环境中聚糖结构基序（由凝集素识别）的分布。但是，基于凝集素染色的聚糖受体的多样性以抽象的方式或充其量从末端聚糖链接（例如A2-3或A2-6连接的唾液酸连接的）来表征。关于植物凝集素的聚糖结合特性的丰富结构和生化信息为开发新策略提供了巨大的潜力，以使用这些凝集素来探测超出末端联系的生理聚糖的精细结构细节。基于这些信息，可以设计用于靶向病原体与其宿主聚糖受体之间的相互作用。基于我们以前在该提案中的努力的基础，我们试图制定以新的方式利用植物凝集素的策略，以获取生理聚糖受体的更详细的结构表征，并开发这些凝集素来靶向病原体 - 宿主聚糖受体相互作用。 公共卫生相关性：复杂的聚糖在生物表面上特别是细胞间表面的界面，在这些界面中，它们在介导细胞相互作用（例如宿主细胞 - 病原体相互作用）中起着关键作用。 在弥合病原体的生理聚糖受体结合与其发病机理和宿主适应的生物学方面存在挑战。这些挑战是由于在适当的生理环境（例如特定细胞和组织表面）中表征聚糖受体的精细结构多样性的局限性。在该提案中，我们试图通过利用植物凝集素的尚未开发的潜力来解决这个问题，以获取在生理表面的聚糖受体的结构特征（例如人类上呼吸道上皮elia）中的结构特征的详细映射，并开发了这些甘眼性作为具有竞争性针对这些受体的工具。