在原核和真核表达系统中生产硒代甲硫氨酸蛋白。

Production of selenomethionyl proteins in prokaryotic and eukaryotic expression systems.

作者信息

Doublié Sylvie

机构信息

Department of Microbiology and Molecular Genetics, University of Vermont, Burlington, VT, USA.

出版信息

Methods Mol Biol. 2007;363:91-108. doi: 10.1007/978-1-59745-209-0_5.

DOI:10.1007/978-1-59745-209-0_5

PMID:17272838

Abstract

The use of selenomethionine as a phasing tool was first reported in 1990. Engineering of selenomethionyl proteins for structure determination is now routine. In fact, selenium is by far the most commonly used anomalous scatterer for multiwavelength anomalous diffraction studies. The past few years have seen new developments, which demonstrated the feasibility of expressing selenomethionyl protein in eukaryotic systems. In this chapter, the different methods available for producing selenomethionine-labeled proteins in bacteria, as well as in yeast and mammalian cells will be presented, along with tips for purifying and crystallizing selenomethionyl proteins.

摘要

1990年首次报道了使用硒代蛋氨酸作为相位工具。如今，为确定结构而对硒代蛋氨酰蛋白质进行工程改造已成为常规操作。事实上，在多波长反常衍射研究中，硒是目前最常用的反常散射体。过去几年出现了新的进展，证明了在真核系统中表达硒代蛋氨酰蛋白质的可行性。在本章中，将介绍在细菌、酵母和哺乳动物细胞中生产硒代蛋氨酸标记蛋白质的不同方法，以及纯化和结晶硒代蛋氨酰蛋白质的技巧。