触发因子协助异二聚体但不是单体荧光素酶的重折叠。

Trigger factor assists the refolding of heterodimeric but not monomeric luciferases.

机构信息

Research Institute for Genetics and Selection of Industrial Microorganisms (GosNIIGenetika), Moscow, 117545, Russia.

出版信息

Biochemistry (Mosc). 2014 Jan;79(1):62-8. doi: 10.1134/S000629791401009X.

Abstract

The refolding of thermally inactivated protein by ATP-independent trigger factor (TF) and ATP-dependent DnaKJE chaperones was comparatively analyzed. Heterodimeric (αβ) bacterial luciferases of Aliivibrio fischeri, Photobacterium leiognathi, and Vibrio harveyi as well as monomeric luciferases of Vibrio harveyi and Luciola mingrelica (firefly) were used as substrates. In the presence of TF, thermally inactivated heterodimeric bacterial luciferases refold, while monomeric luciferases do not refold. These observations were made both in vivo (Escherichia coli ΔdnaKJ containing plasmids with tig gene) and in vitro (purified TF). Unlike TF, the DnaKJE chaperone system refolds both monomeric and heterodimeric luciferases with equal efficiency.

摘要

我们比较了热失活蛋白在非依赖 ATP 的触发因子（TF）和依赖 ATP 的 DnaKJE 伴侣协助下的重折叠情况。我们使用了来自发光杆菌属（Aliivibrio fischeri）、发光杆菌属（Photobacterium leiognathi）和哈维氏弧菌（Vibrio harveyi）的异源二聚体（αβ）细菌荧光素酶，以及哈维氏弧菌和小光萤属（ Luciola mingrelica）（萤火虫）的单体荧光素酶作为底物。在 TF 的存在下，热失活的异源二聚体细菌荧光素酶可以重折叠，而单体荧光素酶则不能。这些观察结果无论是在体内（含有 tig 基因质粒的大肠杆菌ΔdnaKJ）还是在体外（纯化的 TF）都得到了证实。与 TF 不同的是，DnaKJE 伴侣系统能够以相同的效率重折叠单体和异源二聚体荧光素酶。