点击化学在神经可塑性研究中的应用
祁羡杰,阮杭泽,高娟妹,郭 霞,沈万华*
(杭州师范大学浙江省器官发育与再生技术研究重点实验室,杭州 310036)

摘 要:摘 要:神经环路结构和功能的可塑性过程需要大量的新生蛋白参与,鉴定这些蛋白质的组分和在体动态变化无疑会对神经可塑性机制研究提供重要线索。近几年新发展起来的两种点击化学技术,BONCAT ( 生物正交非天然氨基酸标记) 和FUNCAT ( 荧光非天然氨基酸标记) 为神经可塑性研究带来了新的技术手段,它们不仅可以提高检测新生蛋白成分的灵敏度,还可以实时跟踪定位亚细胞新生蛋白的表达情况,已经逐步发展为除荧光和同位素标记蛋白外的另一种重要技术。现就最近几年利用点击化学标记新生蛋白的新技术,包括和质谱联用、筛选新生蛋白以及在各模式动物研究中的应用做一综述。

Click chemistry in neural plasticity
QI Xian-Jie, RUAN Hang-Ze, GAO Juan-Mei, GUO Xia, SHEN Wan-Hua*
(Zhejiang Key Laboratory of Organ Development and Regeneration, Hangzhou Normal University, Hangzhou 310036, China)

Abstract: Abstract: Newly-synthesized proteins are required for neural plasticity and structural plasticity in central nervous system. It will be very helpful to demonstrate the mechanism for neural plasticity by identifying and elucidating the newly-synthesized proteins in specific subcellular compartments. In recent several years, the non-canonical amino acid labeling techniques of BONCAT (bioorthogonal non-canonical amino acid tagging) and FUNCAT (fluorescent non-canonical amino acid tagging) have been widely used to observe protein dynamics and subcellular localization, which provides novel techniques for studying neural plasticity. The next generation of these biological tools not only can increase the sensitivity for detection of newly-synthesized proteins, but also can track the protein dynamics in subcellular compartments in vivo. It has been considered as one of the important techniques for studying newlysynthesized proteins besides the classical methods of fluorescent and radioisotope labeling of proteins. The new techniques for labeling of newly-synthesized proteins by click chemistry, identification of proteomes combined with mass spectrometry (MS) and its applications in animal models are reviewed.

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