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Production and purification of recombinant human interleukin-6 secreted by the yeast Saccharomyces cerevisiae

Yves Guisez, Bernard Tison, Joël Vandekerckhove (UGent) , Jan Demolder (UGent) , Guy Bauw (UGent) , Guy Haegeman (UGent) , Walter Fiers (UGent) and Roland Contreras (UGent)
(1991) EUROPEAN JOURNAL OF BIOCHEMISTRY. 198(1). p.217-222
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Abstract
The coding region of the human interleukin-6 (hIL6) gene was fused to the prepro secretion signal of the alpha-mating factor gene in several yeast host strains. It was found that the KEX-2 protease was unable to cleave the prepro-Lys-Arg-Pro-IL6 sequence, but that unspecific cleavage of the precursor protein had occurred. The prepro-Lys-Arg-Ala-Pro-IL6 sequence, however, was correctly recognized and cleaved by the KEX-2 protease, and IL6 was efficiently secreted into the culture medium. The N-terminal Ala-Pro peptide was removed during processing by wild-type yeast strains, but was retained in a ste13 mutant. IL6 as well as the aberrant proteins were not glycosylated. The transformed cells could secrete up to 30-mu-g/ml IL6. The protein was purified from the medium to homogeneity by ion-exchange chromatography and gel filtration, and had a specific activity of about 2 x 10(8) IU/mg in a proliferation assay.
Keywords
DNA, CELL, TRANSFORMATION, PROTEINS, STRUCTURAL GENE, O-GLYCOSYLATION, ESCHERICHIA-COLI, DIPEPTIDYL AMINOPEPTIDASE, AMINO-ACID SEQUENCE, PREPRO-ALPHA-FACTOR

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Chicago
Guisez, Yves, Bernard Tison, Joël Vandekerckhove, Jan Demolder, Guy Bauw, Guy Haegeman, Walter Fiers, and Roland Contreras. 1991. “Production and Purification of Recombinant Human Interleukin-6 Secreted by the Yeast Saccharomyces Cerevisiae.” European Journal of Biochemistry 198 (1): 217–222.
APA
Guisez, Y., Tison, B., Vandekerckhove, J., Demolder, J., Bauw, G., Haegeman, G., Fiers, W., et al. (1991). Production and purification of recombinant human interleukin-6 secreted by the yeast Saccharomyces cerevisiae. EUROPEAN JOURNAL OF BIOCHEMISTRY, 198(1), 217–222.
Vancouver
1.
Guisez Y, Tison B, Vandekerckhove J, Demolder J, Bauw G, Haegeman G, et al. Production and purification of recombinant human interleukin-6 secreted by the yeast Saccharomyces cerevisiae. EUROPEAN JOURNAL OF BIOCHEMISTRY. 1991;198(1):217–22.
MLA
Guisez, Yves, Bernard Tison, Joël Vandekerckhove, et al. “Production and Purification of Recombinant Human Interleukin-6 Secreted by the Yeast Saccharomyces Cerevisiae.” EUROPEAN JOURNAL OF BIOCHEMISTRY 198.1 (1991): 217–222. Print.
@article{221548,
  abstract     = {The coding region of the human interleukin-6 (hIL6) gene was fused to the prepro secretion signal of the alpha-mating factor gene in several yeast host strains. It was found that the KEX-2 protease was unable to cleave the prepro-Lys-Arg-Pro-IL6 sequence, but that unspecific cleavage of the precursor protein had occurred. The prepro-Lys-Arg-Ala-Pro-IL6 sequence, however, was correctly recognized and cleaved by the KEX-2 protease, and IL6 was efficiently secreted into the culture medium. The N-terminal Ala-Pro peptide was removed during processing by wild-type yeast strains, but was retained in a ste13 mutant. IL6 as well as the aberrant proteins were not glycosylated. The transformed cells could secrete up to 30-mu-g/ml IL6. The protein was purified from the medium to homogeneity by ion-exchange chromatography and gel filtration, and had a specific activity of about 2 x 10(8) IU/mg in a proliferation assay.},
  author       = {Guisez, Yves and Tison, Bernard and Vandekerckhove, Jo{\"e}l and Demolder, Jan and Bauw, Guy and Haegeman, Guy and Fiers, Walter and Contreras, Roland},
  issn         = {0014-2956},
  journal      = {EUROPEAN JOURNAL OF BIOCHEMISTRY},
  keyword      = {DNA,CELL,TRANSFORMATION,PROTEINS,STRUCTURAL GENE,O-GLYCOSYLATION,ESCHERICHIA-COLI,DIPEPTIDYL AMINOPEPTIDASE,AMINO-ACID SEQUENCE,PREPRO-ALPHA-FACTOR},
  language     = {eng},
  number       = {1},
  pages        = {217--222},
  title        = {Production and purification of recombinant human interleukin-6 secreted by the yeast Saccharomyces cerevisiae},
  volume       = {198},
  year         = {1991},
}