Single protein omission reconstitution studies of tetracycline binding to the 30S subunit of Escherichia coli ribosomes
- Univ. of Pennsylvania, Philadelphia (USA)
In previous work the authors showed that on photolysis of Escherichia coli ribosomes in the presence of ({sup 3}H)tetracycline (TC) the major protein labeled is S7, and they presented strong evidence that such labeling takes place from a high-affinity site related to the inhibitory action of TC. In this work they use single protein omission reconstitution (SPORE) experiments to identify those proteins that are important for high-affinity TC binding to the 30S subunit, as measured by both cosedimentation and filter binding assays. With respect to both sedimentation coefficients and relative Phe-tRNA{sup Phe} binding, the properties of the SPORE particles they obtain parallel very closely those measured earlier, with the exception of the SPORE particle lacking S13. A total of five proteins, S3, S7, S8, S14, and S19, are shown to be important for TC binding, with the largest effects seen on omission of proteins S7 and S14. Determination of the protein compositions of the corresponding SPORE particles demonstrates that the observed effects are, for the most part, directly attributable to the omission of the given protein rather than reflecting an indirect effect of omitting one protein on the uptake of another. A large body of evidence supports the notion that four of these proteins, S3, S7, S14, and S19, are included, along with 16S rRNA bases 920-1,396, in one of the major domains of the 30S subunit. The results support the conclusion that the structure of this domain is important for the binding of TC and that, within this domain, TC binds directly to S7.
- OSTI ID:
- 5393748
- Journal Information:
- Biochemistry; (United States), Vol. 29:22; ISSN 0006-2960
- Country of Publication:
- United States
- Language:
- English
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MEMBRANE PROTEINS
BIOCHEMISTRY
RIBOSOMES
CROSS-LINKING
TETRACYCLINES
ESCHERICHIA COLI
PHOTOLYSIS
RECEPTORS
TRITIUM COMPOUNDS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
BACTERIA
CELL CONSTITUENTS
CHEMICAL REACTIONS
CHEMISTRY
DECOMPOSITION
DRUGS
HYDROGEN COMPOUNDS
MICROORGANISMS
ORGANIC COMPOUNDS
PHOTOCHEMICAL REACTIONS
POLYMERIZATION
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550201* - Biochemistry- Tracer Techniques