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http://dx.doi.org/10.5012/bkcs.2007.28.9.1549

Ubiquitin Fusion System for Recombinant Peptide Expression and Purification: Application to the Cytoplasmic Domain of Syndecan-4  

Chae, Young-Kee (Department of Chemistry and Recombinant Protein Expression Center, Sejong University)
Lee, Ha-Yan (Department of Chemistry and Recombinant Protein Expression Center, Sejong University)
Lee, Weon-Tae (Department of Biochemistry and HTSD-NMR & Application NRL, Yonsei University)
Publication Information
Bulletin of the Korean Chemical Society / v.28, no.9, 2007 , pp. 1549-1552 More about this Journal
Abstract
The cytoplasmic domain of syndecan-4, a type I transmembrane heparan sulfate proteoglycan, was overexpressed as a fused form with the ubiquitin molecule in Escherichia coli, and the fusion protein was purified using immobilized metal affinity chromatography (IMAC). The cytoplasmic domain was released from its fusion partner by using yeast ubiquitin hydrolase (YUH), and subsequently purified by reverse phase chromatography. The integrity of the resulting peptide fragment was checked by MALDI-TOF and NMR spectroscopy. The yield of the peptide was 3.0-1.5 mg per liter in LB or minimal medium, respectively. The recombinant expression and purification of this domain will enable us its structural and functional studies using multidimensional NMR spectroscopy.
Keywords
Ubiquitin fusion; Syndecan-4; MALDI-TOF; NMR;
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