Signal peptidase I overproduction results in increased efficiencies of export and maturation of hybrid secretory proteins in Escherichia coli |
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Authors: | Jan Maarten van Dijl Anne de Jong Hilde Smith Sierd Bron and Gerard Venema |
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Institution: | (1) Department of Genetics, Centre of Biological Sciences, NL-9751 NN Haren (Gn), The Netherlands |
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Abstract: | Summary The effects of 25-fold overproduction ofEscherichia coli signal peptidase I (SPase I) on the processing kinetics of various (hybrid) secretory proteins, comprising fusions between
signal sequence functions selected from theBacillus subtilis chromosome and the mature part of TEM-β-lactamase, were studied inE. coli. One precursor (preA2d]-β-lactamase) showed an enhanced processing rate, and consequently, a highly improved release of
the mature enzyme into the periplasm. A minor fraction of a second hybrid precursor (preAl3i]-β-lactamase), which was not
processed under standard conditions of SPase I synthesis, was shown to be processed under conditions of SPase I overproduction.
However, this did not result in efficient release of the mature β-lactamase into the periplasm. In contrast, the processing
rates of wild-type pre-β-lactamase and pre(A2)-β-lactamase, already high under standard conditions, were not detectably altered
by SPase I overproduction. These results demonstrate that the availability of SPase I can be a limiting factor in protein
export inE. coli, in particular with respect to (hybrid) precursor proteins showing low (SPase I) processing efficiencies. |
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Keywords: | Precursor conformation Protein export Signal peptidase I TEM-β -lactamase |
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