Single protein omission reconstitution studies of tetracycline binding to the 30S subunit of Escherichia coli ribosomes

Journal Article · Tue Jun 05 04:00:00 EDT 1990 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00474a024· OSTI ID:5393748

Buck, M; Cooperman, B S ^[1]

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1. 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), Journal Name: Biochemistry; (United States) Vol. 29:22; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
BACTERIA
BIOCHEMISTRY
CELL CONSTITUENTS
CHEMICAL REACTIONS
CHEMISTRY
CROSS-LINKING
DECOMPOSITION
DRUGS
ESCHERICHIA COLI
HYDROGEN COMPOUNDS
MEMBRANE PROTEINS
MICROORGANISMS
ORGANIC COMPOUNDS
PHOTOCHEMICAL REACTIONS
PHOTOLYSIS
POLYMERIZATION
PROTEINS
RECEPTORS
RIBOSOMES
TETRACYCLINES
TRITIUM COMPOUNDS