Expression and purification of recombinant vascular endothelialstatin

We have recently discovered VE-statin (egfl7), a novel protein specifically expressed by endothelial cells during mouse development and in the adult. VE-statin is expected to play an essential role during the formation of new blood vessels. Here, we have expressed the coding sequence for VE-statin i...

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Bibliographic Details
Published in:Protein expression and purification 2006, Vol.46, p.136-142
Main Authors: Caetano, Bertrand, Soncin, Fabrice, Drobecq, Hervé
Format: Article
Language:English
Subjects:
Cancer
Life Sciences
Online Access:Get full text
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Summary:We have recently discovered VE-statin (egfl7), a novel protein specifically expressed by endothelial cells during mouse development and in the adult. VE-statin is expected to play an essential role during the formation of new blood vessels. Here, we have expressed the coding sequence for VE-statin in bacteria and designed a protocol for the production and purification of large amounts of the protein. The protocol is based on an affinity purification step followed by a strong cation-exchange chromatography and a refolding procedure adapted to epidermal growth factor domains and also to the high tendency of the protein to precipitate in concentrated solutions. The purified protein is pure as assessed by SDS-PAGE, immunoblot and mass spectrometry, and is biologically active as a repressor of human aortic smooth muscle cell migration induced by platelet-derived growth factor-BB.
ISSN:1046-5928
1096-0279