Proteins are used by cells as tools to carry out most of their biological processes, ranging from the respiration or metabolism of nutrients, to the sending and processing of intra- and intercellular signals, and to maintaining cellular shape, ensuring growth and controlling cell division.
The University of Cincinnati Drug Discovery Center (UC DDC) has a physical library of 300,000 chemicals that have the general characteristics (i.e. size, chemical properties) of drug-like molecules, and knowledge regarding this library was made available to SAM. With the atomic coordinates of the target protein available, the next step was to predict the inhibitory potential of each of 1,200,000 chemical states derived from the library against PfMAP2. In principle, this can be done using a variety of methods;
Monash University (MU) expressed PfMAP2 in several batches and sent the protein to UC DDC for assay development. It turned out that PfMAP2 was not very active against the substrate that was usually used in kinase assay kits and, to obtain activities sufficiently high for automated high-throughput assays, very high concentrations of protein and substrate were needed.