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PMID: 6423382 Published · ppublish English Journal Article

The binding of 6-demethylchlortetracycline to 70S, 50S and 30S ribosomal particles: a quantitative study by fluorescence anisotropy.

The EMBO journal ·Vol. 3 ·No. 1 ·1984-01-00 ·Pages 121-6

Epe B, Woolley P

Abstract

The binding of demeclocycline (6-demethylchlortetracycline) to ribosomes and ribosomal subunits from Escherichia coli was investigated by using the fluorescence anisotropy of the antibiotic to determine the extent of binding. Binding data obtained from 70S and 30S particles differed fundamentally from those obtained from 50S subunits: the first two showed a strong, specific interaction while the third did not. In addition, all three particles possessed weak, unspecific binding sites. Computer-aided least-squares analysis of the data yielded the following numbers of sites and equilibrium constants: for 30S, n1 = 1, K1 = 2.2 X 10(6) M-1, n2 K2 = 0.029 X 10(6) M-1; for 50S, n1 = 0, n2 K2 = 0.035 X 10(6) M-1; for 70S, n1 = 1, K1 = 3.2 X 10(6) M-1, n2 K2 = 0.082 X 10(6) M-1. These data resolve current disagreement in the literature and are a prerequisite for quantitative studies of the mechanism of inhibition by tetracycline of protein biosynthesis.

MeSH Terms
Computers Demeclocycline/pharmacology Escherichia coli/metabolism Fluorescence Polarization Kinetics Ribosomes/drug effects,metabolism,ultrastructure
Chemicals
Demeclocycline
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Epe B
Woolley P
References (21)
21 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1984-01-00
Pages
121-6
Language
English
Region
England
NLM ID
8208664
PMCID
PMC557307
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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