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dc.contributor.author
Mattle, Daniel
dc.contributor.author
Zeltina, Antra
dc.contributor.author
Woo, Jae-Sung
dc.contributor.author
Goetz, Birke A.
dc.contributor.author
Locher, Kaspar P.
dc.date.accessioned
2021-03-01T07:46:11Z
dc.date.available
2017-06-09T08:22:25Z
dc.date.available
2021-03-01T07:46:11Z
dc.date.issued
2010-11-26
dc.identifier.issn
0022-2836
dc.identifier.issn
1089-8638
dc.identifier.other
10.1016/j.jmb.2010.09.005
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/27158
dc.description.abstract
The periplasmic binding protein HmuT from Yersinia pestis (YpHmuT) is a component of the heme uptake locus hmu and delivers bound hemin to the inner-membrane-localized, ATP-binding cassette (ABC) transporter HmuUV for translocation into the cytoplasm. The mechanism of this process, heme transport across the inner membrane of pathogenic bacteria, is currently insufficiently understood at the molecular level. Here we describe the crystal structures of the substrate-free and heme-bound states of YpHmuT, revealing two lobes with a central binding cleft. Superposition of the apo and holo states reveals a minor tilting motion of the lobes surrounding concomitant with heme binding. Unexpectedly, YpHmuT binds two stacked hemes in a central binding cleft that is larger than those of the homologous periplasmic heme-binding proteins ShuT and PhuT, both of which bind only one heme. The hemes bound to YpHmuT are coordinated via a tyrosine side chain that contacts the Fe atom of one heme and a histidine that contacts the Fe atom of the other heme. The coordinating histidine is only conserved in a subset of periplasmic heme binding proteins suggesting that its presence predicts the ability to bind two heme molecules simultaneously. The structural data are supported by spectroscopic binding studies performed in solution, where up to two hemes can bind to YpHmuT. Isothermal titration calorimetry suggests that the two hemes are bound in discrete, sequential steps and with dissociation constants (KD) of ∼ 0.29 and ∼ 29 nM, which is similar to the affinities observed in other bacterial substrate binding proteins. Our findings suggest that the cognate ABC transporter HmuUV may simultaneously translocate two hemes per reaction cycle.
en_US
dc.language.iso
en
en_US
dc.publisher
Elsevier
en_US
dc.subject
Heme binding
en_US
dc.subject
Iron uptake
en_US
dc.subject
Heme transport protein
en_US
dc.subject
Type III periplasmic binding protein
en_US
dc.subject
Protein crystallography
en_US
dc.title
Two stacked heme molecules in the binding pocket of the periplasmic heme-binding protein HmuT from Yersinia pestis
en_US
dc.type
Journal Article
dc.date.published
2010-09-30
ethz.journal.title
Journal of Molecular Biology
ethz.journal.volume
404
en_US
ethz.journal.issue
2
en_US
ethz.journal.abbreviated
J. Mol. Biol.
ethz.pages.start
220
en_US
ethz.pages.end
231
en_US
ethz.identifier.wos
ethz.identifier.nebis
000049425
ethz.publication.place
Amsterdam
en_US
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02521 - Inst. f. Molekularbiologie u. Biophysik / Inst. Molecular Biology and Biophysics::03652 - Locher, Kaspar / Locher, Kaspar
en_US
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02521 - Inst. f. Molekularbiologie u. Biophysik / Inst. Molecular Biology and Biophysics::03652 - Locher, Kaspar / Locher, Kaspar
ethz.date.deposited
2017-06-09T08:22:54Z
ethz.source
ECIT
ethz.identifier.importid
imp59364d744f47295806
ethz.ecitpid
pub:45940
ethz.eth
yes
en_US
ethz.availability
Metadata only
en_US
ethz.rosetta.installDate
2017-07-15T10:46:54Z
ethz.rosetta.lastUpdated
2021-02-14T06:50:02Z
ethz.rosetta.exportRequired
true
ethz.rosetta.versionExported
true
ethz.COinS
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