Identification of proteins important for tetracycline (TC) binding to ribosomes by single protein omission reconstitution (SPORE) experiments
Abstract
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.
- Authors:
- Publication Date:
- Research Org.:
- Univ. of Pennsylvania, Philadelphia
- OSTI Identifier:
- 6074881
- Report Number(s):
- CONF-870644-
Journal ID: CODEN: FEPRA; TRN: 87-037122
- Resource Type:
- Conference
- Journal Name:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
- Additional Journal Information:
- Journal Volume: 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
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 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
Citation Formats
Buck, M, and Cooperman, B S. Identification of proteins important for tetracycline (TC) binding to ribosomes by single protein omission reconstitution (SPORE) experiments. United States: N. p., 1987.
Web.
Buck, M, & Cooperman, B S. Identification of proteins important for tetracycline (TC) binding to ribosomes by single protein omission reconstitution (SPORE) experiments. United States.
Buck, M, and Cooperman, B S. 1987.
"Identification of proteins important for tetracycline (TC) binding to ribosomes by single protein omission reconstitution (SPORE) experiments". United States.
@article{osti_6074881,
title = {Identification of proteins important for tetracycline (TC) binding to ribosomes by single protein omission reconstitution (SPORE) experiments},
author = {Buck, M and Cooperman, B S},
abstractNote = {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.},
doi = {},
url = {https://www.osti.gov/biblio/6074881},
journal = {Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)},
number = ,
volume = 46:6,
place = {United States},
year = {Fri May 01 00:00:00 EDT 1987},
month = {Fri May 01 00:00:00 EDT 1987}
}