Expression and purification of the anticomplementary peptide Sh-CRIT-ed1 (formerly Sh-TOR-ed1) as a tetramultimer in Escherichia coli
- Authors
- Oh, Kwang-Seok; Na, Do-Kyun; Kweon, Mee-Hyang; Sung, Ha-Chin
- Issue Date
- Feb-2003
- Publisher
- ACADEMIC PRESS INC ELSEVIER SCIENCE
- Keywords
- Sh-CRIT; anticomplementary peptide; expression; repetitive artificial peptide and Escherichia coli
- Citation
- PROTEIN EXPRESSION AND PURIFICATION, v.27, no.2, pp 202 - 209
- Pages
- 8
- Journal Title
- PROTEIN EXPRESSION AND PURIFICATION
- Volume
- 27
- Number
- 2
- Start Page
- 202
- End Page
- 209
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/43530
- DOI
- 10.1016/S1046-5928(02)00598-3
- ISSN
- 1046-5928
1096-0279
- Abstract
- Many complement inhibitors found in plants and other organisms have been recognized as an antiinflammatory drug. Sh-CRIT-ed1 is a complement inhibitory peptide, present on the Schistosoma parasite surface. In the present study, we expressed chemically synthesized oligonucleotides encoding Sh-CRIT-ed1 with an additional hexahistidine codon at the C-terminus and purified in Escherichia coli BL21. The cloned gene, which was multimerized four times in pBlue-script II KS (+) at the isoschizomer sites (BamHI, Bg/II), was named Sh4, and expressed in E. coli BL21 harboring pGEX-KG. The fusion protein (GST-Sh4) was purified with high yield successively by affinity chromatographies of glutathione-Sepharose 4B and Ni-NTA-agarose. Recombinant Sh-CRIT-ed1 I was obtained readily by thrombin digestion and CNBr cleavage of GST-Sh4, and the yield was 9.03 mg from 1-liter culture of E. coli BL21 harboring pGEX-Sh4. The recombinant Sh-CRIT-ed1 showed strong anticomplementary activity (IC50 = 6.02 muM) by complement haemolysis assay. (C) 2002 Elsevier Science (USA). All rights reserved.
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