van Ulsen Peter, Kuhn Karsten, Prinz Thorsten, Legner Harald, Schmid Peter, Baumann Christian, Tommassen Jan
Department of Molecular Microbiology, Utrecht University, Utrecht, The Netherlands.
Proteomics. 2009 Apr;9(7):1771-81. doi: 10.1002/pmic.200800642.
Isobaric labeling reagents such as Tandem Mass Tags (TMT(R)) enable the genome-wide quantification of protein expression levels under different conditions using a gel-free MS/MS-based approach. Here, we applied a TMTduplex approach with two isobaric tags to study the response of the human pathogen Neisseria meningitidis to deprivation of iron, a condition met in the human body. In total, 609 proteins were identified in samples of three independent growth experiments, in which we compared cultures grown in the presence and absence of iron. Expression of 35 proteins was found to be induced or repressed under iron-limiting conditions, including 11 proteins whose ORFs were not previously identified in DNA array studies as being regulated by iron availability at the transcriptional level. These 11 proteins include proteins likely involved in iron metabolism.
等压标记试剂,如串联质谱标签(TMT®),能够使用基于无凝胶MS/MS的方法在不同条件下对全基因组范围内的蛋白质表达水平进行定量。在此,我们应用了一种带有两种等压标签的TMT双链体方法,来研究人类病原体脑膜炎奈瑟菌对铁缺乏(一种在人体中会遇到的情况)的反应。在三个独立生长实验的样本中,总共鉴定出609种蛋白质,在这些实验中我们比较了在有铁和无铁条件下培养的细菌。发现在铁限制条件下35种蛋白质的表达被诱导或抑制,其中包括11种蛋白质,其开放阅读框(ORF)在之前的DNA阵列研究中未被鉴定为在转录水平受铁可用性调控。这11种蛋白质包括可能参与铁代谢的蛋白质。