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Formation of active inclusion bodies in the periplasm of Escherichia coli
Author(s) -
Arié JeanPhilippe,
Miot Marika,
Sassoon Nathalie,
Betton JeanMichel
Publication year - 2006
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.2006.05394.x
Subject(s) - periplasmic space , maltose binding protein , escherichia coli , biology , biochemistry , protein folding , inclusion bodies , fusion protein , folding (dsp implementation) , alkaline phosphatase , phosphatase , enzyme , biophysics , microbiology and biotechnology , recombinant dna , gene , engineering , electrical engineering
Summary To examine the relationship between folding and aggregation in the periplasm of Escherichia coli , we have analysed the cellular fates of exported proteins fused to either the wild‐type maltose‐binding protein (MalE) or the aggregation‐prone variant MalE31. The propensity of fusion proteins to aggregate in the periplasm was determined by the intrinsic folding characteristics of the upstream protein. When β‐lactamase or alkaline phosphatase was linked to the C‐terminus of MalE31, the resultant fusion proteins accumulated in an insoluble form, but retained their catalytic activity. In addition, these protein aggregates induced an extracytoplasmic stress response, similar to unfused MalE31. However, using a fluorescent substrate, we found that alkaline phosphatase activity was present inside periplasmic aggregates. These results suggest that periplasmic inclusion body formation may result in intermolecular interactions between participating proteins without loss of function of the fused enzymes.