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Title: Ribosomal targets for antibiotic drug discovery

Abstract

The present invention relates to methods to identify molecules that binds in the neomycin binding pocket of a bacterial ribosome using structures of an intact bacterial ribosome that reveal how the ribosome binds tRNA in two functionally distinct states, determined by x-ray crystallography. One state positions tRNA in the peptidyl-tRNA binding site. The second, a fully rotated state, is stabilized by ribosome recycling factor (RRF) and binds tRNA in a highly bent conformation in a hybrid peptidyl/exit (P/E) site. Additionally, the invention relates to various assays, including single-molecule assay for ribosome recycling, and methods to identify compounds that interfere with ribosomal function by detecting newly identified intermediate FRET states using known and novel FRET pairs on the ribosome. The invention also provides vectors and compositions with an N-terminally tagged S13 protein.

Inventors:
; ; ; ; ; ;
Issue Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1324693
Patent Number(s):
9441263
Application Number:
14/111,947
Assignee:
The Regents of the University of California (Oakland, CA)
Patent Classifications (CPCs):
G - PHYSICS G01 - MEASURING G01N - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
DOE Contract Number:  
AC03-76SF00098
Resource Type:
Patent
Resource Relation:
Patent File Date: 2012 Apr 14
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES

Citation Formats

Blanchard, Scott C., Feldman, Michael Brian, Wang, Leyi, Doudna Cate, James H., Pulk, Arto, Altman, Roger B., and Wasserman, Michael R. Ribosomal targets for antibiotic drug discovery. United States: N. p., 2016. Web.
Blanchard, Scott C., Feldman, Michael Brian, Wang, Leyi, Doudna Cate, James H., Pulk, Arto, Altman, Roger B., & Wasserman, Michael R. Ribosomal targets for antibiotic drug discovery. United States.
Blanchard, Scott C., Feldman, Michael Brian, Wang, Leyi, Doudna Cate, James H., Pulk, Arto, Altman, Roger B., and Wasserman, Michael R. Tue . "Ribosomal targets for antibiotic drug discovery". United States. https://www.osti.gov/servlets/purl/1324693.
@article{osti_1324693,
title = {Ribosomal targets for antibiotic drug discovery},
author = {Blanchard, Scott C. and Feldman, Michael Brian and Wang, Leyi and Doudna Cate, James H. and Pulk, Arto and Altman, Roger B. and Wasserman, Michael R},
abstractNote = {The present invention relates to methods to identify molecules that binds in the neomycin binding pocket of a bacterial ribosome using structures of an intact bacterial ribosome that reveal how the ribosome binds tRNA in two functionally distinct states, determined by x-ray crystallography. One state positions tRNA in the peptidyl-tRNA binding site. The second, a fully rotated state, is stabilized by ribosome recycling factor (RRF) and binds tRNA in a highly bent conformation in a hybrid peptidyl/exit (P/E) site. Additionally, the invention relates to various assays, including single-molecule assay for ribosome recycling, and methods to identify compounds that interfere with ribosomal function by detecting newly identified intermediate FRET states using known and novel FRET pairs on the ribosome. The invention also provides vectors and compositions with an N-terminally tagged S13 protein.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2016},
month = {9}
}

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Works referenced in this record:

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