MCP：定量蛋白质组学揭示TBP转录因子复合物的动力学机制
生物谷报道：定量蛋白质组学技术目前成为最为热门的技术手段之一。由荷兰乌特勒支大学药理科学系科学家Heck AJ等人刚刚在Mol Cell Proteomics杂志上发表的文章报道，采用定量蛋白质组技术揭示了TATA结合蛋白（TBP）转录因子复合物的动力学调节规律。采用了亲和纯化和isotope蛋白质标签（isotope labeling ）技术结合进行研究。这两种技术结合能有效地，且特异性检测蛋白质之间的相互作用，而且可以排除各类非特异性结合，以减少假阳性。同时还能检测到那些瞬间结合起反应的蛋白质复合物，从而发现以往不能发现的蛋白质之间相互作用，尤其是瞬间结合的反应。在研究细胞周期的同步化中，发现一种TAFs家族的BTAF1交换在其中起到关键性作用，它瞬间与其它蛋白质结构，从而调节了细胞的有丝分裂过程。生物谷网专家认为，这一新技术将在未来得到广泛应用。蛋白质之间相互作用，有时是快速的，甚至瞬间的，很难以检测，传统采用甲醛胶连等技术进行检测，仍然是属于“碰运气”范围。这一新技术不仅更特异性，而且更灵敏，极大拓展人类对蛋白质间相互作用的认识。
英文原文：
Affinity purification in combination with isotope labeling of proteins has proven to be a powerful method to discriminate specific from non-specific interactors. However, in the standard SILAC (Stable isotope labeling by amino acids in cell culture) approach dynamic components may easily be assigned as non-specific. We compared two affinity purification protocols, which in combination revealed information on the dynamics of protein complexes. We focused on the central component in eukaryotic transcription, the human TATA binding protein (TBP), which is involved in different complexes. All known TBP associated factors (TAFs) were detected as specific interactors. Interestingly, one of them, BTAF1, exchanged significantly in cell extracts during the affinity purification. The other TAFs did not display this behavior. Cell cycle synchronization showed that BTAF1 exchange was regulated during mitosis. The combination of the two affinity purification protocols allows a quantitative approach to identify transient components in any protein complex.
出处：
Quantitative proteomics reveals regulation of dynamic components within TATA-binding protein (TBP) transcription complexes.Mol Cell Proteomics. 2007 Dec 17
Mousson F, Kolkman A, Pijnappel WW, Timmers HT, Heck AJ.
Pharmmaceutical Sciences, Utrecht University, Utrecht 3584 CA.
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