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

Autor: Bikmullin AG; Kazan Federal University, Kazan, 420008, Russia. aydar.bikmullin@gmail.com., Nurullina LI; Kazan Federal University, Kazan, 420008, Russia., Garaeva NS; Kazan Federal University, Kazan, 420008, Russia., Klochkova EA; Kazan Federal University, Kazan, 420008, Russia., Blokhin DS; Kazan Federal University, Kazan, 420008, Russia., Golubev AA; Kazan Federal University, Kazan, 420008, Russia.; Institute of Genetics and Molecular and Cellular Biology (IGBMC), Illkirch-Graffenstaden, 67400, France., Validov SZ; Kazan Federal University, Kazan, 420008, Russia., Khusainov IS; Kazan Federal University, Kazan, 420008, Russia.; Institute of Genetics and Molecular and Cellular Biology (IGBMC), Illkirch-Graffenstaden, 67400, France.; Max Planck Institute for Biophysics, Frankfurt-am-Main, 60438, Germany., Usachev KS; Kazan Federal University, Kazan, 420008, Russia. konstantin.usachev@kpfu.ru., Yusupov MM; Kazan Federal University, Kazan, 420008, Russia. marat@igbmc.fr.; Institute of Genetics and Molecular and Cellular Biology (IGBMC), Illkirch-Graffenstaden, 67400, France.
Jazyk: angličtina
Zdroj: Biochemistry. Biokhimiia [Biochemistry (Mosc)] 2020 May; Vol. 85 (5), pp. 545-552.
DOI: 10.1134/S000629792005003X
Abstrakt: 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°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.
Databáze: MEDLINE
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