Ksenia N. Chukhontseva, Vadim V. Salnikov, Oleg S. Morenkov, Sergey V. Kostrov and Ilya V. Demidyuk* Pages 221 - 226 ( 6 )
Background: Protealysin, a zinc metalloprotease of Serratia proteamaculans, is the prototype of a new group within the peptidase family M4. Protealysin-like proteases (PLPs) are widely spread in bacteria but are also found in fungi and archaea. The biological functions of PLPs have not been well studied, but published data showed the involvement of enzymes of this group in the interaction of bacteria with higher organisms, and most likely in the pathogenesis. Such functionality requires the release of the proteases from bacterial cells; however, the data on the cellular localization of PLPs are contradictory and no direct data of this kind have been published.
Objective: Here, the protealysin cellular localization was studied for the first time using immunochemical methods.
Methods and Results: We have produced polyclonal rabbit antibodies against the protealysin precursor. The enzyme was evaluated in cells and medium of periodic culture of S. proteamaculans 94 using Western blotting as well as the enzyme localization was analysed by immunoelectron microscopy. It was shown that more than 99% of the enzyme is in a cell-associated form. Protealysin is accumulated in cells as an inactive precursor. It matures only after the release from cells (after their lysis). Immunoelectron microscopy analysis of bacterial cells has revealed no specific localization of protealysin; it was evenly distributed in the cytoplasm.
Conclusion: The data obtained suggest that S. proteamaculans protealysin and supposedly other protealysin-like proteases are not secreted constitutively and their release from bacteria is likely induced by a certain stimulus such as a contact with a eukaryotic cell. This finding is critical for further studies of the involvement of these enzymes in pathogenesis.
Metalloprotease, protealysin, secretion, Serratia proteamaculans, immunoelectron microscopy, Western blotting.
Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, Kazan Federal University, Kazan, Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, Institute of Molecular Genetics, Russian Academy of Sciences, Moscow