X-ray and neutron small-angle scattering analysis of the complex formed by the Met receptor and the Listeria monocytogenes invasion protein InlB.

2.50
Hdl Handle:
http://hdl.handle.net/10033/23992
Title:
X-ray and neutron small-angle scattering analysis of the complex formed by the Met receptor and the Listeria monocytogenes invasion protein InlB.
Authors:
Niemann, Hartmut H; Petoukhov, Maxim V; Härtlein, Michael; Moulin, Martine; Gherardi, Ermanno; Timmins, Peter; Heinz, Dirk W; Svergun, Dmitri I
Abstract:
The Listeria monocytogenes surface protein InlB binds to the extracellular domain of the human receptor tyrosine kinase Met, the product of the c-met proto-oncogene. InlB binding activates the Met receptor, leading to uptake of Listeria into normally nonphagocytic host cells. The N-terminal half of InlB (InlB(321)) is sufficient for Met binding and activation. The complex between this Met-binding domain of InlB and various constructs of the Met ectodomain was characterized by size exclusion chromatography and dynamic light scattering, and structural models were built using small-angle X-ray scattering and small-angle neutron scattering. Although most receptor tyrosine kinase ligands induce receptor dimerization, InlB(321) consistently binds the Met ectodomain with a 1:1 stoichiometry. A construct comprising the Sema and PSI domains of Met, although sufficient to bind the physiological Met ligand hepatocyte growth factor/scatter factor, does not form a complex with InlB(321) in solution, highlighting the importance of Met Ig domains for InlB binding. Small-angle X-ray scattering and small-angle neutron scattering measurements of ligand and receptor, both free and in complex, reveal an elongated shape for the receptor. The four Ig domains form a bent, rather than a fully extended, conformation, and InlB(321) binds to Sema and the first Ig domain of Met, in agreement with the recent crystal structure of a smaller Met fragment in complex with InlB(321). These results call into question whether receptor dimerization is the basic underlying event in InlB(321)-mediated Met activation and demonstrate differences in the mechanisms by which the physiological ligand hepatocyte growth factor/scatter factor and InlB(321) bind and activate the Met receptor.
Affiliation:
Division of Structural Biology, Helmholtz Center for Infection Research, Inhoffenstrasse 7, D-38124 Braunschweig, Germany.
Citation:
X-ray and neutron small-angle scattering analysis of the complex formed by the Met receptor and the Listeria monocytogenes invasion protein InlB. 2008, 377 (2):489-500 J. Mol. Biol.
Journal:
Journal of molecular biology
Issue Date:
21-Mar-2008
URI:
http://hdl.handle.net/10033/23992
DOI:
10.1016/j.jmb.2008.01.027
PubMed ID:
18262542
Type:
Article
Language:
en
ISSN:
1089-8638
Appears in Collections:
Publications from Division of Molekulare Struktur Biologie (MOSB)

Full metadata record

DC FieldValue Language
dc.contributor.authorNiemann, Hartmut H-
dc.contributor.authorPetoukhov, Maxim V-
dc.contributor.authorHärtlein, Michael-
dc.contributor.authorMoulin, Martine-
dc.contributor.authorGherardi, Ermanno-
dc.contributor.authorTimmins, Peter-
dc.contributor.authorHeinz, Dirk W-
dc.contributor.authorSvergun, Dmitri I-
dc.date.accessioned2008-04-22T11:00:11Z-
dc.date.available2008-04-22T11:00:11Z-
dc.date.issued2008-03-21-
dc.identifier.citationX-ray and neutron small-angle scattering analysis of the complex formed by the Met receptor and the Listeria monocytogenes invasion protein InlB. 2008, 377 (2):489-500 J. Mol. Biol.en
dc.identifier.issn1089-8638-
dc.identifier.pmid18262542-
dc.identifier.doi10.1016/j.jmb.2008.01.027-
dc.identifier.urihttp://hdl.handle.net/10033/23992-
dc.description.abstractThe Listeria monocytogenes surface protein InlB binds to the extracellular domain of the human receptor tyrosine kinase Met, the product of the c-met proto-oncogene. InlB binding activates the Met receptor, leading to uptake of Listeria into normally nonphagocytic host cells. The N-terminal half of InlB (InlB(321)) is sufficient for Met binding and activation. The complex between this Met-binding domain of InlB and various constructs of the Met ectodomain was characterized by size exclusion chromatography and dynamic light scattering, and structural models were built using small-angle X-ray scattering and small-angle neutron scattering. Although most receptor tyrosine kinase ligands induce receptor dimerization, InlB(321) consistently binds the Met ectodomain with a 1:1 stoichiometry. A construct comprising the Sema and PSI domains of Met, although sufficient to bind the physiological Met ligand hepatocyte growth factor/scatter factor, does not form a complex with InlB(321) in solution, highlighting the importance of Met Ig domains for InlB binding. Small-angle X-ray scattering and small-angle neutron scattering measurements of ligand and receptor, both free and in complex, reveal an elongated shape for the receptor. The four Ig domains form a bent, rather than a fully extended, conformation, and InlB(321) binds to Sema and the first Ig domain of Met, in agreement with the recent crystal structure of a smaller Met fragment in complex with InlB(321). These results call into question whether receptor dimerization is the basic underlying event in InlB(321)-mediated Met activation and demonstrate differences in the mechanisms by which the physiological ligand hepatocyte growth factor/scatter factor and InlB(321) bind and activate the Met receptor.en
dc.language.isoenen
dc.subject.meshAnimalsen
dc.subject.meshBacterial Proteinsen
dc.subject.meshBinding Sitesen
dc.subject.meshCHO Cellsen
dc.subject.meshCricetinaeen
dc.subject.meshCricetulusen
dc.subject.meshListeria monocytogenesen
dc.subject.meshMembrane Proteinsen
dc.subject.meshModels, Molecularen
dc.subject.meshNeutronsen
dc.subject.meshProtein Bindingen
dc.subject.meshProtein Structure, Quaternaryen
dc.subject.meshRepressor Proteinsen
dc.subject.meshScattering, Small Angleen
dc.subject.meshSolutionsen
dc.subject.meshX-Raysen
dc.titleX-ray and neutron small-angle scattering analysis of the complex formed by the Met receptor and the Listeria monocytogenes invasion protein InlB.en
dc.typeArticleen
dc.contributor.departmentDivision of Structural Biology, Helmholtz Center for Infection Research, Inhoffenstrasse 7, D-38124 Braunschweig, Germany.en
dc.identifier.journalJournal of molecular biologyen

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