Substrate Profiling of New Enzymes for Drug Discovery

DESCRIPTION (provided by applicant): With a dramatic increase in the number of
complete bacterial genomes in the public and private databases, there exists an
outstanding opportunity for the identification of new antibacterial protein
targets. Interestingly, the accumulation of genomic sequences has been
punctuated by the realization that typically greater than one third of any
genome cannot be annotated for function based on sequence similarity. Several
of these "unknowns" have been shown to be genetically validated drug discovery
targets. However, without a defined biochemical activity, unknowns that prove
to be essential for growth and viability are intractable for the purposes of
drug discovery. Biochemical assay of protein function is critical to the
implementation of high-throughput screening to discover lead compounds, to the
determination of a detailed enzyme mechanism, and to the successful
optimization of those leads to tight-binding inhibitors and ultimately to
efficacious drugs. The proposed research aims to break new ground in the
problem of defining general functional assays, and in understanding the
biochemical activities of proteins of unknown function. The experimental basis
for our proposed methodology involves microcalorimetry and substrate libraries.
In this research proposal, we aim to demonstrate the technology by identifying
substrates for enzymes of unknown function from E. coli, determining the enzyme
mechanisms of these proteins, and designing inhibitors based on the mechanisms.
Establishment of the technology proposed here enables a procedure for the
selection of validated enzyme targets and for the identification of novel
antibacterial drug candidates to combat drug resistant pathogens.

PROPOSED COMMERCIAL APPLICATION:
The invention disclosed here provides a method for identifying substrates and inhibitors for new enzyme targets identified through genomic sequencing. As applied to proteins of unknown function from bacterial genomes, the method of substrate profiling allows one to establish assays for anti-bacterial lead compound discovery in the pharmaceutical industry. The annual world wide market for antibiotic compounds is in excess of $23 billion. Single product sales can exceeed $1 billion. The technology described here enables the discovery of novel antibiotics.