The binding of 6‐demethylchlortetracycline to 70S, 50S and 30S ribosomal particles: a quantitative study by fluorescence anisotropy.
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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.
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The primary (and Tet-1) site is the most highly occupied site in both structures showing tetracycline bound to the 30S subunit, consistent with the high-affinity site being the inhibitory site.
“(i) the primary (and Tet-1) site is the most highly occupied site in both structures showing tetracycline bound to the 30S subunit (4, 37) consistent with the idea that the high-affinity site is the inhibitory site (17, 47, 56).”
From Ribosomal Protection Proteins and Their Mechanism ofTetracyclineResistance (2003) · cites this paper as [17]
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- Authors
- Epe, B., & Woolley, P.
- Affiliations
- University of Würzburg (DE)
- Venue
- The EMBO Journal
- Published
- 1984-01-01
- Volume
- 3
- Issue
- 1
- Pages
- 121-126
- ISSN
- 0261-4189 (ISSN-L)
- 1460-2075
- PMC
- PMC557307
- OpenAlex
- W56361969
- Cited by
- 44
- References
- 21
- Citations by year
- 2025: 1 · 2024: 1 · 2023: 1 · 2022: 2 · 2017: 2 · 2016: 1 · 2015: 2 · 2014: 2 · 2013: 3 · 2012: 2
- FWCI
- 0.46
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- The EMBO Journal (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Interaction Studies and Fluorescence Analysis
- Keywords
- demeclocycline, tetracycline antibiotics, 70S ribosome, 50S ribosomal subunit, 30S ribosomal subunit, Escherichia coli ribosome, binding sites, binding constant, protein synthesis inhibition, fluorescence anisotropy
- MeSH terms
- Computers, Demeclocycline, Escherichia coli, Fluorescence Polarization, Kinetics, Ribosomes
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