DnaK/DnaJ supplementation improves the periplasmic production of human granulocyte-colony stimulating factor in Escherichia coli.
Human granulocyte-colony stimulating factor (hG-CSF) can be efficiently synthesized in E. coli as a fusion to a quimeric signal peptide, but this fusion is only partially exported to the periplasm. We have studied the effects of supplementing the host content in chaperonins on the secretion of prehG-CSF in E. coli. We have found that the DnaK/DnaJ chaperonin system improves the secretion of this recombinant protein. This improvement correlates with an increase in the solubility of the precursor. Only a small fraction of the mature protein is released from the periplasm by an osmotic shock.
Pérez-Pérez
J
J
Departamento de Investigación, Pharmacia Antibióticos Farma S. A., Madrid, Spain.
Martínez-Caja
C
C
Barbero
J L
JL
Gutiérrez
J
J
D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics*; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
C494635:DnaJ protein, E coli; D029968:Escherichia coli Proteins; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; C063842:dnaK protein, E coli