Expression, purification and characterization of soluble human thrombopoietin receptor from Escherichia coli
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- Expression, purification and characterization of soluble human thrombopoietin receptor from Escherichia coli
- Heung Rok Park; Ha Na Im; Young Jun Kang; Myeong Hee Yu; Hyo Jeong Hong
- Bibliographic Citation
- Biotechnology Letters, vol. 22, no. 20, pp. 1611-1617
- Publication Year
- The extracellular domain (edMpl) of human thrombopoietin (TPO) receptor, c-Mpl was expressed in Escherichia coli by changing some nucleotides before and after the translation initiation codon. The mutations increased the expression by approx. 15-fold. The inclusion bodies were solubilized in 8 M guanidine-HCl under reducing conditions and refolded using a glutathione-redox system. The monomeric form of edMpl was purified to near homogeneity by two successive steps of ion-exchange chromatography using DEAE-Sephacel and Mono Q columns. The purified monomeric edMpl inhibited the TPO-dependent cell proliferation, suggesting that it was binding to TPO. Also, antisera raised against the edMpl bound specifically to the soluble receptor secreted by mammalian cells.
- Inclusion bodies; Protein purification; Receptor domain; Thrombopoietin
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- 1. Journal Articles > Journal Articles
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