Sun Na, Tamura Noriko, Tamura Tomohiro, Knispel Roland Wilhelm, Hrabe Thomas, Kofler Christine, Nickell Stephan, Nagy István
Max-Planck-Institute of Biochemistry, D-82152 Martinsried, Germany.
Proteomics. 2009 Jul;9(14):3783-6. doi: 10.1002/pmic.200800892.
We used molecular sieve chromatography in combination with LC-MS/MS to identify protein complexes that can serve as templates in the template matching procedures of visual proteomics approaches. By this method the sample complexity was lowered sufficiently to identify 464 proteins and - on the basis of size distribution and bioinformatics analysis - 189 of them could be assigned as subunits of macromolecular complexes over the size of 300 kDa. From these we purified six stable complexes of Thermoplasma acidophilum whose size and subunit composition - analyzed by electron microscopy and MALDI-TOF-MS, respectively - verified the accuracy of our method.
我们使用分子筛色谱法结合液相色谱-串联质谱法来鉴定蛋白质复合物,这些复合物可作为可视化蛋白质组学方法中模板匹配程序的模板。通过这种方法,样品复杂性被充分降低,从而鉴定出464种蛋白质,并且基于大小分布和生物信息学分析,其中189种可被指定为大小超过300 kDa的大分子复合物的亚基。从中我们纯化了嗜热栖热菌的六种稳定复合物,分别通过电子显微镜和基质辅助激光解吸电离飞行时间质谱分析其大小和亚基组成,验证了我们方法的准确性。