微管蛋白酪氨酸连接酶一步法荧光蛋白标记

One-Step Fluorescent Protein Labeling by Tubulin Tyrosine Ligase.

作者信息

Schumacher Dominik, Leonhardt Heinrich, Hackenberger Christian P R, Helma Jonas

机构信息

Department of Biology II, Ludwig Maximilians Universität München and Center for Integrated Protein Science Munich, Martinsried, Germany.

Department of Chemical-Biology, Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin, Germany.

出版信息

Methods Mol Biol. 2019;2033:167-189. doi: 10.1007/978-1-4939-9654-4_12.

DOI:10.1007/978-1-4939-9654-4_12

PMID:31332754

Abstract

Tub-tag labeling, a novel chemoenzymatic protein functionalization method, facilitates one-step fluorescent labeling of functional biomolecules. The enzyme tubulin tyrosine ligase incorporates coumarin-amino acids to the terminal carboxylic acid of proteins containing a short peptidic recognition sequence called Tub-tag. Here we describe the one-step Tub-tag protein modification protocol in detail and explain its utilization to generate fluorescently labeled proteins for advanced applications in imaging and diagnostics.

摘要

微管标签标记是一种新型的化学酶促蛋白质功能化方法，可实现功能生物分子的一步荧光标记。微管蛋白酪氨酸连接酶将香豆素氨基酸掺入含有短肽识别序列（称为微管标签）的蛋白质的末端羧酸中。在此，我们详细描述了一步微管标签蛋白修饰方案，并解释了其在生成荧光标记蛋白以用于成像和诊断的高级应用中的用途。