Effect of overproduction of heat shock chaperones GroESL and DnaK on human procollagenase production in Escherichia coli.
The effect of overexpression of the heat shock chaperone genes dnaK and groESL on heterologous protein production in Escherichia coli was examined, using a set of related human procollagenase proteins. A diverse range of effects on protein solubility, secretion, and accumulation was observed, and these effects were highly dependent on the particular chaperone/procollagenase pairing involved. Both chaperones caused a large increase in the apparent solubility of a fusion of the LamB signal peptide to procollagenase. GroESL had no effect on the accumulation of mature (secreted) procollagenase, while DnaK suppressed secretion considerably. In the absence of a signal peptide, overexpression of either chaperone resulted in a dramatic increase in both solubility and accumulation of procollagenase. The 10-fold increase in accumulation was associated with an increase in in vivo protein half-life.
Monsanto Corporate Research, Monsanto Company, St. Louis, Missouri 63198.
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D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins / Q000235:genetics*; D018835:Chaperonin 10; D018834:Chaperonin 60; D017364:Collagenases; D003545:Cysteine / Q000378:metabolism; D004792:Enzyme Precursors / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D003012:Microbial Collagenase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D018272:Porins; D021382:Protein Sorting Signals / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D011991:Receptors, Virus / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D018835:Chaperonin 10; D018834:Chaperonin 60; D004792:Enzyme Precursors; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D018272:Porins; D021382:Protein Sorting Signals; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C016532:maltoporins; D017364:Collagenases; C022440:procollagenase; D003012:Microbial Collagenase; C063842:dnaK protein, E coli; D003545:Cysteine