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Abstract
<p class="first" id="P1">Hypertonicity stimulates Nuclear Factor of Activated T-cells
5 (NFAT5) nuclear localization
and transactivating activity. Many transcription factors are known to contain intrinsically
disordered regions (IDRs) which become more structured with local environmental changes
such as osmolality, temperature and tonicity. The transactivating domain of NFAT5
is predicted to be intrinsically disordered under normal tonicity, and under high
NaCl, the activity of this domain is increased. To study the binding of co-regulatory
proteins at IDRs a cDNA construct expressing the NFAT5 TAD was created and transformed
into
<i>E.coli</i> cells. Transformed
<i>E.coli</i> cells were mass produced by fermentation and extracted by cell lysis
to release the
NFAT5 TAD. The NFAT5 TAD was subsequently purified using a His-tag column, cation
exchange chromatography as well as hydrophobic interaction chromatography and then
characterized by mass spectrometry (MS).
</p>