1. [Untitled]
- Author
-
Myeong Hee Yu, Hyo Jeong Hong, Heung Rok Park, Ha Na Im, and Young Jun Kang
- Subjects
Thrombopoietin receptor ,food and beverages ,Bioengineering ,General Medicine ,Biology ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,Molecular biology ,Inclusion bodies ,law.invention ,Biochemistry ,law ,Protein purification ,medicine ,Recombinant DNA ,Extracellular ,Receptor ,Escherichia coli ,Thrombopoietin ,Biotechnology - Abstract
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.
- Published
- 2000