Enzymatic Synthesis of Inositol Pyrophosphates

The availability of well-defined and pure PP-InsP substrates in high quantities is pivotal to study their molecular mechanisms by means of biochemical or biophysical approaches. Here, we present a workflow employing enzymatic synthesis that allows for the quick and easy synthesis and isolation of mammalian PP-InsP isomers. With the respective enzymes, inositol pyrophosphates can be generated in a single reaction step. Isolation and purification of the desired products does not require special laboratory equipment and can be achieved by precipitation of PP-InsPs as magnesium salts. Having access to PP-InsPs will aid the field in discovering and characterizing their molecular mechanisms and binding modes.