Expression and purification of soluble human APRIL in Escherichia coli using ELP-SUMO tag
| Autor: | Shuang Quan Zhang, Lei Ma, Jie Zhang |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Recombinant Fusion Proteins
medicine.medical_treatment SUMO-1 Protein Tumor Necrosis Factor Ligand Superfamily Member 13 Context (language use) Biology medicine.disease_cause Mass Spectrometry law.invention Jurkat Cells Affinity chromatography law Escherichia coli medicine Humans Cloning Molecular Protease Molecular mass Fusion protein Molecular biology Elastin Solubility Biochemistry Recombinant DNA Target protein Peptides Biotechnology |
| Zdroj: | Protein Expression and Purification. 95:177-181 |
| ISSN: | 1046-5928 |
| Popis: | APRIL is a member of the tumor necrosis factor (TNF) family of ligands that mediate tumor cells proliferation as well as survival, depending on the cellular context. In this report, we present a novel method to obtain soluble human APRIL in Escherichia coli using the elastin-like polypeptide and SUMO (ELP-SUMO) tags. The fusion protein with ELP-SUMO tag was expressed in a soluble form at 15 °C. After purification based on inverse transition cycling (ITC) method, the purified ELP-SUMO–hAPRIL fusion protein was subsequently cleaved by SUMO protease to release mature hAPRIL. Following affinity chromatography, the target protein was re-purified with high purity. Finally, about 4.8 mg recombinant hAPRIL was obtained from 1 l bacterial culture with no less than 85% purity. The molecular mass (Mr) of the recombinant hAPRIL was confirmed by MALDI-TOF MS as Mr 16,314. The purified hAPRIL exhibits biological activity on Jurkat cells. It is the first report on soluble production of hAPRIL in E. coli using ELP-SUMO tag. |
| Databáze: | OpenAIRE |
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