Ubiquitin fusion system for recombinant peptide expression and purification: Application to the cytoplasmic domain of syndecan-4

Kee Chae Young, Hayan Lee, Weontae Lee

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1549-1552
Number of pages4
JournalBulletin of the Korean Chemical Society
Volume28
Issue number9
DOIs
Publication statusPublished - 2007 Sep 20

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Syndecan-4
Ubiquitin
Nuclear magnetic resonance spectroscopy
Purification
Fusion reactions
Affinity chromatography
Heparan Sulfate Proteoglycans
Peptides
Peptide Fragments
Hydrolases
Chromatography
Yeast
Escherichia coli
Metals
Molecules
Proteins

All Science Journal Classification (ASJC) codes

  • Chemistry(all)

Cite this

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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.",
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Ubiquitin fusion system for recombinant peptide expression and purification : Application to the cytoplasmic domain of syndecan-4. / Young, Kee Chae; Lee, Hayan; Lee, Weontae.

In: Bulletin of the Korean Chemical Society, Vol. 28, No. 9, 20.09.2007, p. 1549-1552.

Research output: Contribution to journalArticle

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