RAMPED-UP NMR: multiplexed NMR-based screening for drug discovery.
The crucial step in drug discovery is the identification of a lead compound from a vast chemical library by any number of screening techniques. NMR-based screening has the advantage of directly detecting binding of a compound to the target. The spectra resulting from these screens can also be very complex and difficult to analyze, making this an inefficient process. We present here a method, RAMPED-UP NMR, (Rapid Analysis and Multiplexing of Experimentally Discriminated Uniquely Labeled Proteins using NMR) which generates simple spectra which are easy to interpret and allows several proteins to be screened simultaneously. In this method, the proteins to be screened are uniquely labeled with one amino acid type. There are several benefits derived from this unique labeling strategy: the spectra are greatly simplified, resonances that are most likely to be affected by binding are the only ones observed, and peaks that yield little or no information upon binding are eliminated, allowing the analysis of multiple proteins easily and simultaneously. We demonstrate the ability of three different proteins to be analyzed simultaneously for binding to two different ligands. This method will have significant impact in the use of NMR spectroscopy for both the lead generation and lead optimization phases of drug discovery by its ability to increase screening throughput and the ability to examine selectivity. To the best of our knowledge, this is the first time in any format that multiple proteins can be screened in one tube.
Zartler
Edward R
ER
Discovery Chemistry Research and Technologies, Biology Research Technologies and Proteins, Lilly Research Labs, Lilly Corporate Center, Eli Lilly & Co., Indianapolis, Indiana 46285, USA.
Hanson
Jeffrey
J
Jones
Bryan E
BE
Kline
Allen D
AD
Martin
George
G
Mo
Huaping
H
Shapiro
Michael J
MJ
Wang
Rong
R
Wu
Haiping
H
Yan
Jiangli
J
D020650:Combinatorial Chemistry Techniques / Q000379:methods; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry
D011506:Proteins