Sun Y P, Andrews P C, Smith D L
Department of Medicinal Chemistry and Pharmacognosy, Purdue University, West Lafayette, Indiana 47907.
J Protein Chem. 1990 Apr;9(2):151-7. doi: 10.1007/BF01025306.
A new procedure to selectively identify disulfide-containing peptides in extracts of biological tissues is described. Disulfide-containing peptides are detected by their UV absorbance and electrochemical (EC) activity after chromatographic separation, and subsequently identified by fast atom bombardment mass spectrometry (FABMS). This combination of fractionation by HPLC and selective detection is attractive because it is rapid, highly specific for disulfide-containing peptides, and applicable to all disulfide-containing peptides that may be present in complex biological mixtures. Useful procedures for applying the method are demonstrated with tissue extracts from bovine pituitary and catfish pancreas. In addition to finding the expected disulfide-containing peptides, evidence for two forms of catfish insulin are presented. The merits of this and other methods used to detect peptides in similar tissue extracts are discussed.
本文描述了一种用于选择性鉴定生物组织提取物中含二硫键肽段的新方法。含二硫键的肽段在色谱分离后通过紫外吸收和电化学（EC）活性进行检测，随后通过快原子轰击质谱（FABMS）进行鉴定。高效液相色谱（HPLC）分级分离与选择性检测相结合的方法具有吸引力，因为它快速、对含二硫键的肽段具有高度特异性，并且适用于复杂生物混合物中可能存在的所有含二硫键的肽段。利用牛垂体和鲶鱼胰腺的组织提取物展示了应用该方法的实用步骤。除了找到预期的含二硫键肽段外，还提供了两种鲶鱼胰岛素形式的证据。讨论了该方法以及用于检测类似组织提取物中肽段的其他方法的优点。
