Identification of proteins important for tetracycline (TC) binding to ribosomes by single protein omission reconstitution (SPORE) experiments
TC inhibits protein synthesis in E. coli by interfering with aminoacyl-tRNA binding to the ribosomal A site, and there is strong evidence that such inhibition results from TC binding to a high affinity site on the 30S subunit. The SPORE approach has been used to define those 30S proteins that are potentially important for high affinity TC binding, measured as the (/sup 3/H)-TC co-sedimenting with the reconstitution particle through a sucrose density gradient. Reverse phase-HPLC has been used both to prepare ribosomal proteins and to analyze the protein content of reconstituted particles. The results obtained so far show that protein S7, as well as some proteins linked to S7 in the 30S assembly map, are important for TC binding, whereas other ribosomal proteins are not. These results are in very good accord with their earlier photoaffinity labeling studies that strongly implicated S7 as forming part of the TC binding site. Interestingly, protein S18, which is photolabeled by TC to a high extent but in a non-site specific manner, appears to be unimportant for TC binding.
- Research Organization:
- Univ. of Pennsylvania, Philadelphia
- OSTI ID:
- 6074881
- Report Number(s):
- CONF-870644-; TRN: 87-037122
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
PROTEINS
BIOSYNTHESIS
TETRACYCLINES
BIOCHEMICAL REACTION KINETICS
RECEPTORS
AFFINITY
ESCHERICHIA COLI
INHIBITION
LIQUID COLUMN CHROMATOGRAPHY
RIBOSOMES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
BACTERIA
CELL CONSTITUENTS
CHROMATOGRAPHY
DRUGS
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MEMBRANE PROTEINS
MICROORGANISMS
ORGANIC COMPOUNDS
ORGANOIDS
REACTION KINETICS
SEPARATION PROCESSES
SYNTHESIS
550201* - Biochemistry- Tracer Techniques