Preparation and characterization of human interleukin-5 expressed in recombinant Escherichia coli.
The gene coding for human interleukin-5 was synthesized and expressed in Escherichia coli under control of a heat-inducible promoter. High-level expression, 10-15% of total cellular protein, was achieved in E. coli. The protein was produced in an insoluble state. A simple extraction, renaturation and purification scheme is described. The recombinant protein was found to be a homodimer, similar to the natural murine-derived protein. Despite the lack of glycosylation, high specific activities were obtained in three 'in vitro' biological assays. Physical characterization of the protein showed it to be mostly alpha-helical, supporting the hypothesis that a conformational similarity exists among certain cytokines.
Glaxo Institute for Molecular Biology S.A., Geneva, Switzerland.
A E
AE
Proudfoot
D
D
Fattah
E H
EH
Kawashima
A
A
Bernard
P T
PT
Wingfield
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002454:Cell Differentiation; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004220:Disulfides / Q000032:analysis; D004563:Electrochemistry; D004804:Eosinophils / Q000166:cytology; D004926:Escherichia coli / Q000378:metabolism*; D015870:Gene Expression; D006358:Hot Temperature; D006801:Humans; D007231:Infant, Newborn; D015848:Interleukin-5 / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D046911:Macromolecular Substances; D051379:Mice; D008970:Molecular Weight; D011401:Promoter Regions, Genetic / Q000235:genetics; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D012995:Solubility; D013438:Sulfhydryl Compounds / Q000032:analysis
D000596:Amino Acids; D004220:Disulfides; D015848:Interleukin-5; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D013438:Sulfhydryl Compounds