Affinity purification of ribosomes with a lethal G2655C mutation in 23 S rRNA 
that affects the translocation

Leonov, Andrei A.; Sergiev, Petr V.; Bodganov, Alexej A.; Brimacombe, Richard; Dontsova, Olga A.

doi:10.1074/jbc.M302873200

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Affinity purification of ribosomes with a lethal G2655C mutation in 23 S rRNA that affects the translocation

Leonov, A. A., Sergiev, P. V., Bodganov, A. A., Brimacombe, R., & Dontsova, O. A. (2003). Affinity purification of ribosomes with a lethal G2655C mutation in 23 S rRNA that affects the translocation. Journal of Biological Chemistry, 278(28), 25664-25670. doi:10.1074/jbc.M302873200.

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Item Permalink: https://hdl.handle.net/11858/00-001M-0000-0010-89F9-A Version Permalink: https://hdl.handle.net/11858/00-001M-0000-0010-89FA-8

Genre: Journal Article

Alternative Title : J. Biol. Chem.

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Creators:
Leonov, Andrei A., Author
Sergiev, Petr V., Author
Bodganov, Alexej A.¹, Author
Brimacombe, Richard², Author
Dontsova, Olga A., Author

Affiliations:
1Max Planck Society, ou_persistent13
2Ribosomes, Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433558

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Abstract: A method for preparation of Escherichia coli ribosomes carrying lethal mutations in 23 S rRNA was developed. The method is based on the site-directed incorporation of a streptavidin binding tag into functionally neutral sites of the 23 S rRNA and subsequent affinity chromatography. It was tested with ribosomes mutated at the 23 S rRNA position 2655 (the elongation factor (EF)-G binding site). Ribosomes carrying the lethal G2655C mutation were purified and studied in vitro. It was found in particular that this mutation confers strong inhibition of the translocation process but only moderately affects GTPase activity and binding of EF-G.

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Language(s): eng - English

Dates: Date issued: 2003-07-11

Publication Status: Issued

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Identifiers: eDoc: 174591
ISI: 000183920200050
DOI: 10.1074/jbc.M302873200

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Title: Journal of Biological Chemistry

Alternative Title : J. Biol. Chem.

Source Genre: Journal

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Pages: - Volume / Issue: 278 (28) Sequence Number: - Start / End Page: 25664 - 25670 Identifier: ISSN: 0021-9258