Expression and assembly of active human cardiac troponin in Escherichia coli

Cardiomyopathy-related mutations in human cardiac troponin subunits, including troponin C (hcTnC), troponin I (hcTnI), and troponin T (hcTnT), are well-documented. Recently, it has been recognised that human cardiac troponin (hcTn) is a sophisticated allosteric system. Therefore, the effect of drugs on this protein complex should be studied with assembled hcTn rather than a short fragment of a subunit or the subunit itself. Here, we describe the expression and assembly of active hcTn in Escherichia coli, a novel method that is rapid and simple, and produces large amounts of functional hcTn.

► High yield hcTroponin assembly in Escherichia coli has been made possible.
► Only two columns are necessary to obtain a large amount of purified hcTroponin.
► Transformation and purification of hcTroponin can be performed within one week.
► High-throughput screening of cardiac disease mutations is now feasible.