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@ARTICLE{VanderMeeren:141270,
author = {Van der Meeren, R. and Wen, Y. and Van Gelder, P. and
Tommassen, J. and Devreese, B. and Savvides, S. N. and DESY},
title = {{N}ew insights into the assembly of bacterial secretins:
structural studies of the periplasmic domain of {X}cp{Q}
from {P}seudomonas aeruginosa},
journal = {The journal of biological chemistry},
volume = {288},
issn = {0021-9258},
address = {Bethesda, Md.},
publisher = {Soc.},
reportid = {PHPPUBDB-25356},
pages = {1214-1225},
year = {2013},
abstract = {The type II secretion system is a multiprotein assembly
spanning the inner and outer membranes in Gram-negative
bacteria. It is found in almost all pathogenic bacteria
where it contributes to virulence, host tissue colonization,
and infection. The exoproteins are secreted across the outer
membrane via a large translocation channel, the secretin,
which typically adopts a dodecameric structure. These
secretin channels have large periplasmic N-terminal domains
that reach out into the periplasm for communication with the
inner membrane platform and with a pseudopilus structure
that spans the periplasm. Here we report the crystal
structure of the N-terminal periplasmic domain of the
secretin XcpQ from Pseudomonas aeruginosa, revealing a
two-lobe dimeric assembly featuring parallel subunits
engaging in well defined interactions at the tips of each
lobe. We have employed structure-based engineering of
disulfide bridges and native mass spectrometry to show that
the periplasmic domain of XcpQ dimerizes in a
concentration-dependent manner. Validation of these insights
in the context of cellular full-length XcpQ and further
evaluation of the functionality of disulfide-linked XcpQ
establishes that the basic oligomerization unit of XcpQ is a
dimer. This is consistent with the notion that the
dodecameric secretin assembles as a hexamer of dimers to
ensure correct projection of the N-terminal domains into the
periplasm. Therefore, our studies provide a key conceptual
advancement in understanding the assembly principles and
dynamic function of type II secretion system secretins and
challenge recent studies reporting monomers as the basic
subunit of the secretin oligomer.},
keywords = {Amino Acid Sequence / Bacterial Proteins: chemistry /
Bacterial Proteins: genetics / Bacterial Proteins:
metabolism / Electrophoresis, Polyacrylamide Gel / Mass
Spectrometry / Molecular Sequence Data / Periplasm:
metabolism / Protein Conformation / Pseudomonas aeruginosa:
metabolism / Recombinant Proteins: chemistry / Recombinant
Proteins: genetics / Recombinant Proteins: metabolism /
Secretin: metabolism / Sequence Homology, Amino Acid /
Bacterial Proteins (NLM Chemicals) / Recombinant Proteins
(NLM Chemicals) / Secretin (NLM Chemicals)},
cin = {EMBL},
ddc = {570},
cid = {$I:(DE-H253)EMBL_-2012_-20130307$},
pnm = {DORIS Beamline D1.2 (POF2-54G13)},
pid = {G:(DE-H253)POF2-D1.2-20130405},
experiment = {EXP:(DE-H253)D-D1.2-20150101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:23188826},
pmc = {pmc:PMC3543004},
UT = {WOS:000313570300042},
doi = {10.1074/jbc.M112.432096},
url = {https://bib-pubdb1.desy.de/record/141270},
}