Azizi Amedeo A, Kang Sung-Ung, Freilinger Angelika, Gruber-Olipitz Mariella, Chen Wei-Qiang, Yang Jae-Won, Hengstschläger Markus, Slavc Irene, Lubec Gert
Department of Pediatrics and Adolescent Medicine, Medical University of Vienna, Wahringer Gurtel 18-20, Vienna, Austria.
J Proteome Res. 2008 Aug;7(8):3412-22. doi: 10.1021/pr800149p. Epub 2008 Jun 27.
Systematic work on differential protein expression in mitosis is limited, and we therefore used neuroblastoma cells (N1E-115) incubated with either colcemid or nocodazole to arrest mitosis. Proteins were identified by MALDI-TOF/TOF and nano-LC-ESI-MS/MS with subsequent quantification of spot volumes with specific software. Immunoblotting was used for verification of selected proteins. Levels of 10 individual proteins were increased and levels of 6 proteins were decreased concordantly by both treatments. These proteins were constituents of heat shock and chaperone, cytoskeleton, proteasomal, heterochromatin, and DNA replication signaling as well as housekeeping and metabolic systems. Identification of mitosis-dependent proteins is of importance for the interpretation of previous work and for designing future experiments.
关于有丝分裂中差异蛋白质表达的系统性研究有限,因此我们使用用秋水仙酰胺或诺考达唑孵育的神经母细胞瘤细胞(N1E-115)来阻滞有丝分裂。通过基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF/TOF)和纳升级液相色谱-电喷雾串联质谱(nano-LC-ESI-MS/MS)鉴定蛋白质,随后使用特定软件对斑点体积进行定量。免疫印迹法用于验证选定的蛋白质。两种处理均使10种蛋白质的水平升高,6种蛋白质的水平一致降低。这些蛋白质是热休克和伴侣蛋白、细胞骨架、蛋白酶体、异染色质、DNA复制信号以及管家和代谢系统的组成部分。鉴定有丝分裂依赖性蛋白质对于解释先前的工作和设计未来的实验具有重要意义。