A fusion protein consisting of human calmodulin (CaM) and glutathione
S-transferase (GST) was
produced by gene fusion. The fusion protein was overexpressed in
Escherichia coli as a soluble form
and purified with one-step affinity chromatography using
glutathione−Sepharose. The protein had
the modulating activity of CaM and the binding capability to
glutathione of GST. Phosphodiesterase,
which is a CaM dependent enzyme, was activated by the fusion protein,
with the Ca2+ level equal to
the level equivalent to a native CaM. Furthermore, CaM could be
immobilized on a solid-phase matrix
through the use of GST moiety while its modulating activity was
retained. Phosphodiesterase activity
was switched on and off by the immobilized CaM with or without
Ca2+, and repeated use of CaM was
demonstrated.
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