Wang Sheng, Zhang Luyan, Yang Pengyuan, Chen Gang
School of Pharmacy and Department of Chemistry, Fudan University, Shanghai, China.
Proteomics. 2008 Jul;8(13):2579-82. doi: 10.1002/pmic.200800086.
In this report, infrared (IR) radiation was employed to enhance the efficiency of tryptic proteolysis for peptide mapping. Protein solutions containing trypsin in sealed transparent Eppendorf tubes were allowed to digest under an IR lamp at 37 degrees C. The feasibility and performance of the novel proteolysis approach were demonstrated by the digestion of BSA and myoglobin (MYO) and the digestion time was significantly reduced to 5 min. The obtained digests were identified by MALDI-TOF MS with the sequence coverages of 69% (BSA) and 90% (MYO) that were much better than those obtained by conventional in-solution tryptic digestion. The present IR-assisted proteolysis strategy is simple and efficient, offering great promise for high-throughput protein identification.
在本报告中,采用红外(IR)辐射来提高用于肽图谱分析的胰蛋白酶解效率。将含有胰蛋白酶的蛋白质溶液置于密封的透明艾本德管中,在37℃的红外灯下进行消化。通过对牛血清白蛋白(BSA)和肌红蛋白(MYO)的消化证明了这种新型蛋白酶解方法的可行性和性能,消化时间显著缩短至5分钟。通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)对所得消化产物进行鉴定,序列覆盖率分别为69%(BSA)和90%(MYO),远优于传统溶液内胰蛋白酶消化所获得的结果。目前这种红外辅助蛋白酶解策略简单高效,为高通量蛋白质鉴定提供了广阔前景。