MASS MAPPING TO SHOW DISULFIDE BOND CONNECTIVY IN PROTEINS W/ ADJACENT CYSTEINES

质量图谱显示蛋白质与相邻半胱氨酸的二硫键连接性

• 批准号: 6258785
• 负责人: JACK T WATSON
• 金额: $ 0.34万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-01 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6258785
• 关键词: biological products; biomedical equipment development; biomedical resource; biotechnology; proteins; technology /technique development

Conventional approaches to disulfide bond mapping rely on proteases to cleave the peptide backbone between all cysteine residues. Because proteases cannot cleave between adjacent cysteines, proteins containing this structural feature are refractory to the conventional approach. We have demonstrated that our cyanylation methodology can cleave the adjacent cysteine residues in long R3 insulin-like growth factor (LR3IGF), an 83-residue protein containing six cysteines in the form of three disulfide bonds. Analysis of the cleavage products of two singly reduced isoforms of LR3IGF by LC-MS using electrospray ionization verified that cleavage between Cys60 and Cys61 had been achieved. The results of this study also proved that the disulfide connectivity in LR3IGF-1 is homologous to that in insulin-like growth factor (IGF-1).

二硫键作图的常规方法依赖于 蛋白酶切割所有半胱氨酸之间的肽骨架 残基 因为蛋白酶不能在相邻的半胱氨酸之间切割， 含有这种结构特征的蛋白质是难治的， 传统的方法。 我们已经证明了我们的氰化 方法可以切割长R3中的相邻半胱氨酸残基 胰岛素样生长因子（LR 3 IGF），一种含有83个残基的蛋白质， 六个半胱氨酸以三个二硫键的形式存在。 分析 LC-MS法测定LR 3 IGF的两种单一还原亚型的裂解产物 使用电喷雾离子化证实Cys 60和 Cys 61已达到。 这项研究的结果也证明， LR 3 IGF-1中的二硫键连接与 胰岛素样生长因子（IGF-1）。