Expression of the Soluble Extracellular Domain of Human Thrombopoietin Receptor Using a Maltose-Binding Protein-Affinity Fusion System
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Accession number;04A0108798
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| Title;Expression of the Soluble Extracellular Domain of Human Thrombopoietin Receptor Using a Maltose-Binding Protein-Affinity Fusion System |
| Author;
ZHANG Q
(Zhongshan Univ., Guangzhou, Chn)
PAN R-M
(Zhongshan Univ., Guangzhou, Chn)
GE Y-C
(Zhongshan Univ., Guangzhou, Chn)
XU P
(Zhongshan Univ., Guangzhou, Chn)
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Journal Title;Biol Pharm Bull
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Journal Code:S0989A
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ISSN:0918-6158
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VOL.27;NO.2;PAGE.219-221(2004)
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| Figure&Table&Reference;FIG.4, REF.9 |
| Pub. Country;Japan |
| Language;English |
| Abstract;The thrombopoietin (TPO) receptor (Mpl) belongs to the family of ligand-dependent cytokine receptors and plays a functional role in regulating platelet production. The signaling capacity largely depends on the binding of TPO to the extracellular domains of the TPO receptor (Mpl-EC). Because the expression level of Mpl in human tissue is very low, studies on the functional and spatial characteristics of its ligand-binding sites have been limited. In the present study, we report the expression and purification of Mpl-EC as a fusion with the maltose-binding protein (MBP), designated MBP-Mpl-EC. MBP-Mpl-EC was expressed in the cytoplasm of Escherichia coli as a soluble fusion protein. Specific binding of TPO to purified MBP-Mpl-EC was demonstrated by a dot-blot assay and surface plasmon resonance. We conclude that bacterial expression of MBP-Mpl-EC yields large amounts of protein with correct folding and that it can be used for further structure and function analyses. (author abst.) |
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