Michael A Wasserman

US Patent:

20140127682, May 8, 2014

Filed:

Apr 14, 2012

Appl. No.:

14/111947

Inventors:

Scott C. Blanchard - New York NY, US
Michael Brian Feldman - Setauket NY, US
Leyi Wang - New York NY, US
James H. Doudna Cate - Berkeley CA, US
Arto Pulk - Berkeley CA, US
Roger B. Altman - New York NY, US
Michael R. Wasserman - New York NY, US

Assignee:

THE REGENTS OF THE UNIVERSITY OF CALIFORNIA - Oakland CA
CORNELL UNIVERSITY - Ithaca NY

International Classification:

C12Q 1/68

US Classification:

435 61, 530402

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