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（申请人提供）：复杂聚糖普遍分布在细胞间表面的界面上，作为各种微生物病原体的受体，在确定病原体的宿主细胞和组织向性方面发挥着关键作用。尽管它们在宿主-病原体相互作用中很重要，但将聚糖-蛋白质相互作用的生化和生物物理特异性与这些相互作用调节的生物功能联系起来一直具有挑战性。这些挑战源于在生理背景（例如不同组织的上皮表面）中表征聚糖受体的精细结构粒度的困难。因此，需要在组织、细胞和分子水平上桥接聚糖的结构表征。在这些水平上解码生理聚糖受体的多样性不仅有助于从理解发病机制的角度识别聚糖受体靶点，而且还有助于制定对抗病原体感染的治疗策略。我们已经开始通过开发多方面的框架来解码聚糖-蛋白质相互作用的结构-功能关系来解决上述问题。通过结合组织凝集素染色和基于从代表性细胞类型中分离的聚糖的质谱分析，我们在组织和细胞水平上连接聚糖的表征方面迈出了一步。 植物凝集素传统上用于研究聚糖结构基序（由凝集素识别）在细胞和组织表面的生理环境中的分布。然而，基于凝集素染色的聚糖受体的多样性已以抽象方式或最多以末端聚糖连接（例如α2-3或α2-6连接的唾液酸）来表征。关于植物凝集素聚糖结合特性的丰富的结构和生化信息为开发新策略提供了巨大的潜力，利用这些凝集素来探索超越末端连接的生理聚糖的更精细的结构细节。基于这些信息，可以设计凝集素来靶向病原体与其宿主聚糖受体之间的相互作用。基于我们之前在此提案中所做的努力，我们寻求开发以新方式利用植物凝集素的策略，以获得生理聚糖受体更详细的结构特征，并开发这些凝集素以靶向病原体-宿主聚糖受体相互作用。 公共健康相关性：复杂的聚糖装饰生物表面，特别是细胞间表面的界面，它们在介导细胞与细胞相互作用（例如宿主细胞与病原体相互作用）中发挥着关键作用。 将病原体的生理聚糖受体结合与其发病机制和宿主适应的生物学联系起来存在挑战。这些挑战源于在适当的生理环境（例如特定的细胞和组织表面）中表征聚糖受体的精细结构多样性的局限性。在本提案中，我们寻求通过利用植物凝集素未开发的潜力来解决这个问题，以获得生理表面（例如人类上呼吸道上皮细胞）聚糖受体结构特征的细微差别的详细图谱，并将这些凝集素开发为工具竞争性地靶向这些受体。