TRYPTIC DIGESTION OF INTERFERON COMPONENTS

干扰素成分的胰酶消化

• 批准号: 3792473
• 负责人: J BEKISZ
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3792473
• 关键词: endopeptidases; high performance liquid chromatography; interferons; protein structure; proteolysis

Twenty two interferon components which have been purified from Wellferon in our laboratory will be digested using trypsin TPCK. All digestion will be carried out at 37 degrees C for a period of 16 to 18 hours. The digested material will then be passed over a C-18 HPLC column to separate the individual fragments. This will enable us to generate a tryptic digest map (chromatogram) for each component. The fragments from each component will then be collected and assayed for protein sequence and amino acid analysis. To date, we have only been able to partially characterize the components. By sequencing the digestion fragments, we should be able to determine the full protein sequence for each component. The amino acid analysis will provide a compositional evaluation of each component. Characterization of the entire molecule is important if we are to develop hydrophobicity and antigenicity profiles as well as determining the secondary structure of the component.

Wellferon中提纯的22种干扰素成分 在我们实验室将用胰酶TPCK消化。全部消化 将在37摄氏度下进行16至18小时。这个 消化后的物质将通过C-18高效液相色谱柱分离 单独的碎片。这将使我们能够产生胰酶 每种成分的消化图谱(色谱图)。每一张照片的碎片 然后收集成分并进行蛋白质序列分析和 氨基酸分析。到目前为止，我们只能部分地 确定组件的特征。通过对消化片段进行测序，我们 应该能够确定每个成分的完整蛋白质序列。 氨基酸分析将提供每种物质的组成评估 组件。 如果我们要发展，整个分子的特性是很重要的 疏水性和抗原性分布以及测定 零部件的二级结构。