Format

Send to

Choose Destination
Anal Biochem. 2017 Jan 15;517:53-55. doi: 10.1016/j.ab.2016.11.010. Epub 2016 Nov 16.

Removal of protein S1 from Escherichia coli ribosomes without the use of affinity chromatography.

Author information

1
Institute of Protein Research of the Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia.
2
Institute of Protein Research of the Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia. Electronic address: alexch@vega.protres.ru.

Abstract

The paper reports an inexpensive and efficient procedure for the removal of protein S1 from E. coli ribosomes. It comprises incubation of ribosomes in a pyrimidine polyribonucleotide solution followed by centrifugation of the sample through a sucrose cushion. To avoid co-sedimentation of the S1-bound polypyrimidine with the ribosomes, its length should not exceed several hundred nucleotides. Unlike popular affinity chromatography through a poly(U) Sepharose or poly(U) cellulose column, the method tolerates limited polyribonucleotide degradation by eventual traces of ribonucleases, and can readily be incorporated into standard protocols for the isolation of ribosomes by centrifugation.

KEYWORDS:

Centrifugation; E. coli ribosomes; Poly(C); Poly(U); Protein S1

PMID:
27865825
DOI:
10.1016/j.ab.2016.11.010
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center