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@ARTICLE{Kuhle:207429,
author = {Kuhle, K. and Krausze, J. and Curth, U. and Roessle,
Manfred and Heuner, K. and Lang, C. and Flieger, A.},
title = {{O}ligomerization {I}nhibits {L}egionella {P}neumophila
{P}la{B} {P}hospholipase {A} {A}ctivity},
journal = {The journal of biological chemistry},
volume = {289},
number = {27},
issn = {1083-351X},
address = {Bethesda, Md.},
publisher = {Soc.},
reportid = {PUBDB-2015-01339},
pages = {18657 - 18666},
year = {2014},
note = {(c) by The American Society for Biochemistry and Molecular
Biology, Inc. Post referee full text in progress.},
abstract = {The intracellularly replicating lung pathogen Legionella
pneumophila consists of an extraordinary variety of
phospholipases, including at least 15 different
phospholipases A (PLA). Among them, PlaB, the first
characterized member of a novel lipase family, is a
hemolytic virulence factor that exhibits the most prominent
PLA activity in L. pneumophila. We analyzed here protein
oligomerization, the importance of oligomerization for
activity, addressed further essential regions for activity
within the PlaB C terminus, and the significance of
PlaB-derived lipolytic activity for L. pneumophila
intracellular replication. We determined by means of
analytical ultracentrifugation and small angle x-ray
scattering analysis that PlaB forms homodimers and
homotetramers. The C-terminal 5, 10, or 15 amino acids,
although the individual regions contributed to PLA activity,
were not essential for protein tetramerization. Infection of
mouse macrophages with L. pneumophila wild type, plaB
knock-out mutant, and plaB complementing or various mutated
plaB-harboring strains showed that catalytic activity of
PlaB promotes intracellular replication. We observed that
PlaB was most active in the lower nanomolar concentration
range but not at or only at a low level at concentration
above 0.1 $\mu M$ where it exists in a dimer/tetramer
equilibrium. We therefore conclude that PlaB is a virulence
factor that, on the one hand, assembles in inactive
tetramers at micromolar concentrations. On the other hand,
oligomer dissociation at nanomolar concentrations activates
PLA activity. Our data highlight the first example of
concentration-dependent phospholipase inactivation by
tetramerization, which may protect the bacterium from
internal PLA activity, but enzyme dissociation may allow its
activation after export.},
cin = {EMBL / DOOR},
ddc = {570},
cid = {I:(DE-H253)EMBL-20120731 / I:(DE-H253)HAS-User-20120731},
pnm = {DORIS Beamline D1.2 (POF2-54G13) / 6G3 - PETRA III
(POF3-622)},
pid = {G:(DE-H253)POF2-D1.2-20130405 / G:(DE-HGF)POF3-6G3},
experiment = {EXP:(DE-H253)D-D1.2-20150101 / EXP:(DE-H253)P-P12-20150101},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000339062900004},
doi = {10.1074/jbc.M114.573196},
url = {https://bib-pubdb1.desy.de/record/207429},
}