Development of a sensitive method to measure heparan sulfate 6-O-endosulfatase

开发一种灵敏的方法来测量硫酸乙酰肝素 6-O-内切硫酸酯酶

基本信息

  • 批准号:
    10599763
  • 负责人:
  • 金额:
    $ 35万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2023
  • 资助国家:
    美国
  • 起止时间:
    2023-02-01 至 2024-01-31
  • 项目状态:
    已结题

项目摘要

The proposal is aimed to develop a sensitive method to measure the activity of heparan sulfate 6-O-endosulfatase (SULF). SULF is an enzyme that removes the 6-O-sulfo groups from heparan sulfate polysaccharides to modulate the functions of heparan sulfate. Evidence has demonstrated that the level of SULF contributes to the progression of cancers; however, the method that measures the activity from biological sources is lacking. In this phase I SBIR proposal, we propose to use a LC-MS/MS method to determine the activity from plasma and solid tissues. The crucial innovation is that we will use structurally homogeneous heparan sulfate oligosaccharides as substrates to address the issue for the selectivity of the measurement. In addition, we plan to use 13C-labeled oligosaccharide substrates and coupled with disaccharide analysis using a LC-MS/MS, which is expected to increase the sensitivity and add quantitation capability. Aim 1 is designed to find the optimal oligosaccharides for purified SULF enzyme, demonstrating the feasibility of using oligosaccharides to measure the activity of SULFS, including both SULF-1 and SULF-2. We plan to test different oligosaccharides in the size ranging from hexasaccharide (6-mer) to octasaccharide (8- mer). Aim 2 is improve the quantification accuracy using 13C-labeled HS oligosaccharide substrates and LC-MS/MS. Here, we plan to use 13C-labeled oligosaccharide substrate in the analysis. The use of 13C-labeled substrate will eliminate the interference from endogenous HS. Instead of directly measuring de-sulfated oligosaccharide, we plan to degrade the 13C-labeled oligosaccharides into disaccharides using heparin lyases followed by LC-MS/MS analysis. We expect that disaccharide analysis will significantly increase the sensitivity compared to the measurement of intact oligosaccharides, offering a sensitive and quantitative method to measure the activity of SULF from tissues and plasma. In the phase II analysis, we plan to expand the analysis to a wide range of cancer tissues. The success will lead to a necessary tool for glycoscience to investigate the roles of heparan sulfate in cancers.
该建议旨在开发一种灵敏的方法来测量肝素的活性

项目成果

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Guowei Su其他文献

Guowei Su的其他文献

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{{ truncateString('Guowei Su', 18)}}的其他基金

Synthesis of the library of heparan sulfate hexasaccharide mimetics
硫酸乙酰肝素六糖模拟物文库的合成
  • 批准号:
    10759528
  • 财政年份:
    2023
  • 资助金额:
    $ 35万
  • 项目类别:
Developing Heparan Sulfate Glycan Array
开发硫酸乙酰肝素聚糖阵列
  • 批准号:
    10005531
  • 财政年份:
    2017
  • 资助金额:
    $ 35万
  • 项目类别:

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