In vitro Reconstitution of the S. aureus 30S Ribosomal Subunit and RbfA Factor Complex for Structural Studies

被引：4

作者：

Bikmullin, A. G. ^{[1
]}

Nurullina, L. I. ^{[1
]}

Garaeva, N. S. ^{[1
]}

Klochkova, E. A. ^{[1
]}

Blokhin, D. S. ^{[1
]}

Golubev, A. A. ^{[1
,2
]}

Validov, Sh. Z. ^{[1
]}

Khusainov, I. Sh. ^{[1
,2
,3
]}

Usachev, K. S. ^{[1
]}

Yusupov, M. M. ^{[1
,2
]}

机构：

[1] Kazan Fed Univ, Kazan 420008, Russia

[2] Inst Genet & Mol & Cellular Biol IGBMC, F-67400 Illkirch Graffenstaden, France

[3] Max Planck Inst Biophys, D-60438 Frankfurt, Germany

来源：

BIOCHEMISTRY-MOSCOW | 2020年 / 85卷 / 05期

基金：

俄罗斯基础研究基金会;

关键词：

Staphylococcus aureus; ribosome; cryo-electron microscopy; RbfA; translation factor; 30S subunit assembly; CRYOELECTRON MICROSCOPY;

D O I：

10.1134/S000629792005003X

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Ribosome-binding factor A (RbfA) from Staphylococcus aureus is a cold adaptation protein that is required for the growth of pathogenic cells at low temperatures (10-15 degrees C). RbfA is involved in the processing of 16S rRNA, as well as in the assembly and stabilization of the small 30S ribosomal subunit. Structural studies of the 30S-RbfA complex will help to better understand their interaction, the mechanism of such complexes, and the fundamental process such as 30S subunit assembly that determines and controls the overall level of protein biosynthesis. This article describes protocols for preparation of RbfA and the small 30S ribosomal subunits and reconstitution and optimization of the 30S-RbfA complex to obtain samples suitable for cryo-electron microscopy studies.

引用

页码：545 / 552

页数：8

共 27 条

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