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Precise localization of an overproduced periplasmic protein in Escherichia coli: use of double immuno‐gold labelling
Author(s) -
Bernadac A.,
Bolla J. M.,
Lazdunski C.,
Inouye M.,
Pages J. M.
Publication year - 1987
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1111/j.1768-322x.1987.tb00580.x
Subject(s) - periplasmic space , biology , escherichia coli , cytoplasm , subcellular localization , inner membrane , bacterial outer membrane , overproduction , antiserum , secretion , organelle , compartment (ship) , microbiology and biotechnology , biochemistry , enzyme , membrane , antigen , gene , genetics , oceanography , geology
The subcellular localization of beta‐lactamase produced by a secretion‐cloning vector pIN‐III was studied by immunolabelling of frozen thin sections of Escherichia coli. Using double immuno‐gold detections and internal reference proteins, it is shown here that beta‐lactamase encoded by this vector can be exported and that its overproduction leads to aggregation within the periplasm. This aggregation induces the appearance of electron‐dense areas immunolabelled by the antiserum directed against the beta‐lactamase at the external side of the cytoplasmic membrane. The overproduced enzyme is also secreted to the medium in vesicles budding from the outer membrane of lpp strains.