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Structure of the pneumococcal l , d ‐carboxypeptidase DacB and pathophysiological effects of disabled cell wall hydrolases DacA and DacB
Author(s) -
Abdullah Mohammed R.,
GutiérrezFernández Javier,
Pribyl Thomas,
Gisch Nicolas,
Saleh Malek,
Rohde Manfred,
Petruschka Lothar,
Burchhardt Gerhard,
Schwudke Dominik,
Hermoso Juan A.,
Hammerschmidt Sven
Publication year - 2014
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/mmi.12729
Subject(s) - peptidoglycan , carboxypeptidase , biology , mutant , function (biology) , biochemistry , cell , microbiology and biotechnology , enzyme , gene
Summary Bacterial cell wall hydrolases are essential for peptidoglycan turnover and crucial to preserve cell shape. The d , d ‐carboxypeptidase DacA and l , d ‐carboxypeptidase DacB of S treptococcus pneumoniae function in a sequential manner. Here, we determined the structure of the surface‐exposed lipoprotein DacB . The crystal structure of DacB , radically different to that of DacA , contains a mononuclear Zn 2+ catalytic centre located in the middle of a large and fully exposed groove. Two different conformations were found presenting a different arrangement of the active site topology. The critical residues for catalysis and substrate specificity were identified. Loss‐of‐function of DacA and DacB altered the cell shape and this was consistent with a modified peptidoglycan peptide composition in dac mutants. Contrary, an lgt mutant lacking lipoprotein diacylglyceryl transferase activity required for proper lipoprotein maturation retained l , d ‐carboxypeptidase activity and showed an intact murein sacculus. In addition we demonstrated pathophysiological effects of disabled DacA or DacB activities. Real‐time bioimaging of intranasal infected mice indicated a substantial attenuation of Δ dacB and Δ dacA Δ dacB pneumococci, while Δ dacA had no significant effect. In addition, uptake of these mutants by professional phagocytes was enhanced, while the adherence to lung epithelial cells was decreased. Thus, structural and functional studies suggest DacA and DacB as optimal drug targets.