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Title
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High yield expression and NMR characterization of Arkadia E3 ubiquitin ligase RING-H2 finger domain.
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Authors
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N.G.Kandias,
C.T.Chasapis,
D.Bentrop,
V.Episkopou,
G.A.Spyroulias.
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Ref.
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Biochem Biophys Res Commun, 2009,
378,
498-502.
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PubMed id
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Abstract
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E3 ubiquitin ligases play a key role in the recognition of target proteins and
the degradation by 26S proteasomes. Arkadia is the first example of an E3
ubiquitin ligase that positively regulates TGF-beta family signaling. It has
been shown to induce ubiquitin-dependent degradation of negative regulators of
TGF-beta signaling through its C-terminal RING domain. Structural analysis of
Arkadia RING domain is needed to elucidate its enzymatic properties. For such
studies efficient production of pure and correctly folded Arkadia protein is
required. Here we report the recombinant expression in Escherichia coli and
purification of the C-terminal RING domain of Arkadia. NMR analysis of the
soluble construct reveals a stable folded protein suitable for high resolution
structural studies.
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