The binding of ribosomal protein SI to Sl-depleted 30S and 70S ribosomes. A fluorescence anisotropy study of the effects of Mg2+

Dixie J. Goss, Lawrence J Parkhurst, Arkesh M. Mehta, Albert J. Wahba

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Abstract

We have determined the equilibrium constants for the binding of AEDANS-labelled SI to Sl-depleted 30S and 70S ribosomes. For "tight" ribosomes,2+the association of SI increases with the sixth power of Mg concentration, but for 30S subunits and "loose" ribosomes, there is virtually no dependence of the association on Mg2+ over the same concentration range, 2-10 mM in Mg2+. The binding of SI to 70S ribosomes at 10 mM Mg2+ is stabilized by 2 kcal/mol compared to the binding to 30S subunits. When intact SI binds to tight ribosomes, the fluorescence anisotropy is that for virtually complete rotational immobilization. The anisotropies vary considerably with the preparation and treatment of both SI and ribosomes and these variations are detailed here. The results suggest the linkage of Mg2+ -dependent conformational changes in the intact ribosomes, perhaps including rRNA, and the binding of SI.

Original languageEnglish (US)
Pages (from-to)5589-5602
Number of pages14
JournalNucleic acids research
Volume11
Issue number16
DOIs
StatePublished - Aug 25 1983

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Fluorescence Polarization
Ribosomal Proteins
Ribosomes
Fluorescence
Anisotropy
Carrier Proteins
Association reactions
Proteins
Protein
Equilibrium constants
Linkage
Ribosome Subunits
Preparation
Vary
Immobilization
Dependent
Range of data
Mm

ASJC Scopus subject areas

  • Genetics

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The binding of ribosomal protein SI to Sl-depleted 30S and 70S ribosomes. A fluorescence anisotropy study of the effects of Mg2+. / Goss, Dixie J.; Parkhurst, Lawrence J; Mehta, Arkesh M.; Wahba, Albert J.

In: Nucleic acids research, Vol. 11, No. 16, 25.08.1983, p. 5589-5602.

Research output: Contribution to journalArticle

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N2 - We have determined the equilibrium constants for the binding of AEDANS-labelled SI to Sl-depleted 30S and 70S ribosomes. For "tight" ribosomes,2+the association of SI increases with the sixth power of Mg concentration, but for 30S subunits and "loose" ribosomes, there is virtually no dependence of the association on Mg2+ over the same concentration range, 2-10 mM in Mg2+. The binding of SI to 70S ribosomes at 10 mM Mg2+ is stabilized by 2 kcal/mol compared to the binding to 30S subunits. When intact SI binds to tight ribosomes, the fluorescence anisotropy is that for virtually complete rotational immobilization. The anisotropies vary considerably with the preparation and treatment of both SI and ribosomes and these variations are detailed here. The results suggest the linkage of Mg2+ -dependent conformational changes in the intact ribosomes, perhaps including rRNA, and the binding of SI.

AB - We have determined the equilibrium constants for the binding of AEDANS-labelled SI to Sl-depleted 30S and 70S ribosomes. For "tight" ribosomes,2+the association of SI increases with the sixth power of Mg concentration, but for 30S subunits and "loose" ribosomes, there is virtually no dependence of the association on Mg2+ over the same concentration range, 2-10 mM in Mg2+. The binding of SI to 70S ribosomes at 10 mM Mg2+ is stabilized by 2 kcal/mol compared to the binding to 30S subunits. When intact SI binds to tight ribosomes, the fluorescence anisotropy is that for virtually complete rotational immobilization. The anisotropies vary considerably with the preparation and treatment of both SI and ribosomes and these variations are detailed here. The results suggest the linkage of Mg2+ -dependent conformational changes in the intact ribosomes, perhaps including rRNA, and the binding of SI.

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