DEVELOPMENT OF A NEW METHOD FOR ISOLATING THE C-TERMINAL PROTEOLYTIC PEPTIDE

分离 C 端蛋白水解肽的新方法的开发

• 批准号: 6279538
• 负责人: Salvatore Sechi
• 金额: $ 0.84万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-12-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6279538
• 关键词: biomedical resource; proteins; spectrometry; technology /technique

A new approach for the characterization of the C-terniiiials of proteins has been developed. The strategy involves three steps. First, the protein is degraded enzymatically with trypsin or Lys-C. Second the C-tenninal peptide is separated from all the other proteolysis products through the use of anhydrotrypsin, a modified trypsin that binds only peptides containing K or R at the C-terminus. Finally, the C-terminal peptide is characterized by MALDI. The strategy was tested and shown to work well on a series of designed synthetic peptides as well as on the transcription factor Hairy, where the C-terminal peptide was readily isolated and identified.

表征 C 端基的新方法 蛋白质已被开发出来。 该策略包括三个步骤。 首先，用胰蛋白酶或 Lys-C 酶促降解蛋白质。 其次，C 末端肽与所有其他肽分开 通过使用脱水胰蛋白酶（一种改良的 仅结合 C 末端含有 K 或 R 的肽的胰蛋白酶。 最后，C 端肽通过 MALDI 进行表征。 这 该策略经过测试，证明在一系列设计上效果良好 合成肽以及转录因子 Hairy，其中 C 端肽很容易分离和鉴定。