权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Novel Reagents for Monitoring Sulfation Patterns in Heparin and Heparan Sulfate

用于监测肝素和硫酸乙酰肝素硫酸化模式的新型试剂

基本信息

批准号：
9909150
负责人：
Sheng-Cheng Wu
金额：
$ 22.5万
依托单位：
GLYCOSENSORS AND DIAGNOSTICS, LLC
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-01 至 2022-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9909150
关键词：
Adhesions Affinity Affinity Chromatography Antibodies Anticoagulants Antithrombins Binding Binding Proteins Biochemical Reaction Biological Biological Assay Biological Markers Carbohydrates Cell surface Cells Competitive Binding Complex Computer Analysis Computer Simulation Crystallization Detection Directed Molecular Evolution Disease Disease Marker Distal Enzymes Event Future Gene Library Health Heparan Sulfate Biosynthesis Heparin Heparitin Sulfate Heparitin sulfotransferase Histological Techniques Image In Situ Letters Libraries Location Malignant neoplasm of prostate Mediating Monitor Mutagenesis Neoplasm Metastasis Oligonucleotides Oligosaccharides Parents Pathogenicity Pathologic Pathway interactions Pattern Point Mutation Polysaccharides Positioning Attribute Process Property Protein Analysis Protein Isoforms Reagent Reporting Scaffolding Protein Signal Transduction Site Specificity Structure Structure-Activity Relationship Sulfate Techniques Therapeutic Time Tissues Variant Vertebral column Viral Western Blotting Yeasts angiogenesis axon guidance base combinatorial commercial application design disease diagnosis heparan-sulfate 2-sulfotransferase histological stains improved in vivo imaging insight migration mutant neoplastic cell novel pathogen preference receptor response scaffold screening sulfotransferase sulfurtransferase

项目摘要

PROJECT SUMMARY Oligo- and polysaccharides participate in fundamental molecular interactions, and their location on cell surfaces makes them ideal candidates as biomarkers for various disease states. Heparan sulfate (HS) is a highly sulfated linear polysaccharide that participates in an assortment of cellular signaling events that are either advantageous or pathogenic, depending upon the sulfation pattern along the polysaccharide chain. High-affinity binding probes, such as antibodies, are an effective means for discerning molecular interactions within cells; however, few reagents are sufficiently specific for monitoring the sulfation patterns of HS in situ. This project will develop a new class of reagents that is capable of recognizing specific sulfation patterns within the HS chain by exploiting the inherent specificity of enzymes (sulfotransferases) that are responsible for transferring the sulfate groups to the polysaccharide during HS biosynthesis. Protein scaffolds will be created by catalytically inactivating the sulfotransferases, and the affinity of the scaffolds will be enhanced via computationally-guided single point mutations, as well as by directed evolution. By selectively targeting either the substrate or product of the enzymatic reaction, each inactive sulfotransferase has the potential to bind to two specific sequences of HS. This project will focus on sulfotransferases HS 2-O-Sulfotransferase (HS2ST) and 3-O-Sulfotransferase Isoform 1 (HS3ST-1); however, there are numerous isoforms of various enzymes from the biosynthetic pathway of HS that could be targeted in future studies. Both enzymes interact with biologically significant sulfation patterns. For example, HS2ST has been implicated in a variety of functions including angiogenesis, axon guidance, and prostate cancer. The HS3ST-1 enzyme is directly responsible for the anticoagulant properties of heparin, by increasing the affinity of HS for antithrombin. These reagents would be immensely useful for tracking changes in sulfation patterns via typical antibody-mediated assays, including affinity purification, western blotting, in situ histological staining, and in vivo imaging.

项目总结