ABSTRACT
A statistically exhaustive, 8800 compound tripeptidal amidomethylcoumarin library was synthesized as discreet compounds using solid-phase combinatorial chemistry. A subset of the compounds was purified by HPLC and tested in a high-throughput fluorometric assay against several known serine and cysteine proteases to demonstrate the utility of this library for profiling protease substrate specificity.
Subject(s)
Coumarins/chemical synthesis , Endopeptidases/metabolism , Fluorescent Dyes/chemical synthesis , Chromatography, High Pressure Liquid , Combinatorial Chemistry Techniques , Coumarins/chemistry , Fluorescent Dyes/chemistry , Molecular Probes , Substrate SpecificityABSTRACT
We have proposed computer-generated models of the catalytic subunits of the serine-threonine protein phosphatases PP1 and PP2A complexed with their endogenous substrate phospho-DARPP-32, and several known naturally occurring inhibitors. This study is part of an overall effort to elucidate the signal transduction pathways in which PP1 and PP2A may play an important role.
Subject(s)
Enzyme Inhibitors/chemistry , Okadaic Acid/chemistry , Phosphoprotein Phosphatases/antagonists & inhibitors , Amino Acid Sequence , Enzyme Inhibitors/metabolism , Enzyme Inhibitors/pharmacology , Models, Molecular , Molecular Sequence Data , Okadaic Acid/metabolism , Okadaic Acid/pharmacology , Phosphoprotein Phosphatases/chemistry , Phosphoprotein Phosphatases/metabolism , Protein Binding , Sequence Homology, Amino Acid , Structure-Activity RelationshipABSTRACT
The okadaic acid class of naturally occurring toxins is a structurally diverse group of molecules that inhibit the protein phosphatases PP1 and PP2A. Studies providing information about the mode of binding between the toxins and the phosphatases contribute to an overall understanding of the signal transduction pathways in which the phosphatases are involved.
