Mapping disulfide bonds from sub-micrograms of purified proteins or micrograms of complex protein mixtures

• DOI: 10.1007/s41048-018-0050-6
• Publication Date: 2018-04-23T13:54:30Z
• Authors: Shan Lu; Yong Cao; Sheng-Bo Fan; Zhen-Lin Chen; Run-Qian Fang; Si-Min He; Meng-Qiu Dong
• Keywords: 0303 health sciences; 03 medical and health sciences; Protocol

Disulfide bonds are vital for protein functions, but locating the linkage sites has been a challenge in chemistry, especially when quantity of sample is small or complexity high. In 2015, our laboratory developed sensitive and efficient method mapping disulfide from simple complex samples (Lu et al. Nat Methods 12:329, 2015). This based on liquid chromatography-mass spectrometry (LC-MS) powerful data analysis software tool named pLink. To facilitate application this method, we present step-by-step protocols three types samples-purified proteins solution, SDS-PAGE gels, mixtures solution. The minimum amount required can be as low several hundred nanograms purified proteins, tens micrograms mixture hundreds proteins. entire workflow-from preparation to LC-MS analysis-is described great detail. We believe that protocol easily implemented any with access fast-scanning, high-resolution, accurate-mass system.