Crystal structure of an engineered YopM-InlB hybrid protein

Breitsprecher D, Gherardi E, Bleymüller W, Niemann H (2014)
BMC Structural Biology 14(1): 12.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
Download
OA
Autor*in
Breitsprecher, Dennis; Gherardi, Ermanno; Bleymüller, WillemUniBi; Niemann, HartmutUniBi
Abstract / Bemerkung
BACKGROUND: The multi-domain protein InlB (internalin B) from Listeria monocytogenes is an agonist of the human receptor tyrosine kinase MET. Only the internalin domain directly interacts with MET. The internalin domain consists of seven central leucine-rich repeats (LRRs) flanked by an N-terminal helical cap domain and a C-terminal immunoglobulin-like structure. A potential function of the N-terminal cap in receptor binding could so far not be demonstrated by deleting the cap, since the cap is also implicated in nucleating folding of the LRR domain. RESULTS: We generated an InlB variant (YopM-InlB) in which the InlB cap domain was replaced by the unrelated N-terminal capping structure of the LRR protein YopM from Yersinia enterocolitica. The crystal structure of the engineered protein shows that it folds properly. Because the first LRR is structurally closely linked to the cap domain, we exchanged LRR1 along with the cap domain. This resulted in unexpected structural changes extending to LRR2 and LRR3, which are deeply involved in MET binding. As a consequence, the binding of YopM-InlB to MET was substantially weaker than that of wild type InlB. The engineered protein was about one order of magnitude less active in colony scatter assays than wild type InlB. CONCLUSIONS: We obtained a well-behaved InlB variant with an altered N-terminal capping structure through protein design. The reduced affinity for MET precludes a straightforward interpretation of the results from cell-based assays. Still, the engineered hybrid protein induced cell scatter, suggesting that the cap is required for folding and stability of InlB but is not essential for interactions that assemble the signalling-active receptor complex. The cap swap approach described here is clearly applicable to other L. monocytogenes internalins and other LRR proteins such as YopM and may yield useful structure/function correlates within this protein family.
Stichworte
Capping structure; Chimeric protein; Internalin; Leucine-rich repeat; Cap domain; Hybrid protein; Protein stability; LRR; Protein engineering; Protein chimera
Erscheinungsjahr
2014
Zeitschriftentitel
BMC Structural Biology
Band
14
Ausgabe
1
Art.-Nr.
12
ISSN
1472-6807
Finanzierungs-Informationen
Open-Access-Publikationskosten wurden durch die Deutsche Forschungsgemeinschaft und die Universität Bielefeld gefördert.
Page URI
https://pub.uni-bielefeld.de/record/2671451

Zitieren

Breitsprecher D, Gherardi E, Bleymüller W, Niemann H. Crystal structure of an engineered YopM-InlB hybrid protein. BMC Structural Biology. 2014;14(1): 12.
Breitsprecher, D., Gherardi, E., Bleymüller, W., & Niemann, H. (2014). Crystal structure of an engineered YopM-InlB hybrid protein. BMC Structural Biology, 14(1), 12. doi:10.1186/1472-6807-14-12
Breitsprecher, Dennis, Gherardi, Ermanno, Bleymüller, Willem, and Niemann, Hartmut. 2014. “Crystal structure of an engineered YopM-InlB hybrid protein”. BMC Structural Biology 14 (1): 12.
Breitsprecher, D., Gherardi, E., Bleymüller, W., and Niemann, H. (2014). Crystal structure of an engineered YopM-InlB hybrid protein. BMC Structural Biology 14:12.
Breitsprecher, D., et al., 2014. Crystal structure of an engineered YopM-InlB hybrid protein. BMC Structural Biology, 14(1): 12.
D. Breitsprecher, et al., “Crystal structure of an engineered YopM-InlB hybrid protein”, BMC Structural Biology, vol. 14, 2014, : 12.
Breitsprecher, D., Gherardi, E., Bleymüller, W., Niemann, H.: Crystal structure of an engineered YopM-InlB hybrid protein. BMC Structural Biology. 14, : 12 (2014).
Breitsprecher, Dennis, Gherardi, Ermanno, Bleymüller, Willem, and Niemann, Hartmut. “Crystal structure of an engineered YopM-InlB hybrid protein”. BMC Structural Biology 14.1 (2014): 12.
Alle Dateien verfügbar unter der/den folgenden Lizenz(en):
Copyright Statement:
Dieses Objekt ist durch das Urheberrecht und/oder verwandte Schutzrechte geschützt. [...]
Volltext(e)
Access Level
OA Open Access
Zuletzt Hochgeladen
2019-09-06T09:18:22Z
MD5 Prüfsumme
ed1854b494e9551c399be6256e7d718c


1 Zitation in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Unraveling Gardnerella vaginalis Surface Proteins Using Cell Shaving Proteomics.
Marín E, Haesaert A, Padilla L, Adán J, Hernáez ML, Monteoliva L, Gil C., Front Microbiol 9(), 2018
PMID: 29867878

48 References

Daten bereitgestellt von Europe PubMed Central.

Entry of Listeria monocytogenes into hepatocytes requires expression of inIB, a surface protein of the internalin multigene family.
Dramsi S, Biswas I, Maguin E, Braun L, Mastroeni P, Cossart P., Mol. Microbiol. 16(2), 1995
PMID: 7565087
Structural basis of MET receptor dimerization by the bacterial invasion protein InlB and the HGF/SF splice variant NK1
AUTHOR UNKNOWN, 1834
InIB-dependent internalization of Listeria is mediated by the Met receptor tyrosine kinase.
Shen Y, Naujokas M, Park M, Ireton K., Cell 103(3), 2000
PMID: 11081636
Structural insights into Met receptor activation.
Niemann HH., Eur. J. Cell Biol. 90(11), 2011
PMID: 21242015
GW domains of the Listeria monocytogenes invasion protein InlB are required for potentiation of Met activation.
Banerjee M, Copp J, Vuga D, Marino M, Chapman T, van der Geer P, Ghosh P., Mol. Microbiol. 52(1), 2004
PMID: 15049825
InlB: an invasion protein of Listeria monocytogenes with a novel type of surface association.
Braun L, Dramsi S, Dehoux P, Bierne H, Lindahl G, Cossart P., Mol. Microbiol. 25(2), 1997
PMID: 9282740
Multiple regions of internalin B contribute to its ability to turn on the Ras-mitogen-activated protein kinase pathway.
Copp J, Marino M, Banerjee M, Ghosh P, van der Geer P., J. Biol. Chem. 278(10), 2002
PMID: 12488439
Fold and function of the InlB B-repeat.
Ebbes M, Bleymuller WM, Cernescu M, Nolker R, Brutschy B, Niemann HH., J. Biol. Chem. 286(17), 2011
PMID: 21345802
Investigation of the mechanism of binding between internalin B and heparin using surface plasmon resonance.
Hrtska SC, Kemp MM, Munoz EM, Azizad O, Banerjee M, Raposo C, Kumaran J, Ghosh P, Linhardt RJ., Biochemistry 46(10), 2007
PMID: 17305366
Structure of the human receptor tyrosine kinase met in complex with the Listeria invasion protein InlB.
Niemann HH, Jager V, Butler PJ, van den Heuvel J, Schmidt S, Ferraris D, Gherardi E, Heinz DW., Cell 130(2), 2007
PMID: 17662939
The leucine-rich repeat structure.
Bella J, Hindle KL, McEwan PA, Lovell SC., Cell. Mol. Life Sci. 65(15), 2008
PMID: 18408889
Internalins from the human pathogen Listeria monocytogenes combine three distinct folds into a contiguous internalin domain.
Schubert WD, Gobel G, Diepholz M, Darji A, Kloer D, Hain T, Chakraborty T, Wehland J, Domann E, Heinz DW., J. Mol. Biol. 312(4), 2001
PMID: 11575932
Folding and stability of the leucine-rich repeat domain of internalin B from Listeri monocytogenes.
Freiberg A, Machner MP, Pfeil W, Schubert WD, Heinz DW, Seckler R., J. Mol. Biol. 337(2), 2004
PMID: 15003459
Structure of internalin, a major invasion protein of Listeria monocytogenes, in complex with its human receptor E-cadherin.
Schubert WD, Urbanke C, Ziehm T, Beier V, Machner MP, Domann E, Wehland J, Chakraborty T, Heinz DW., Cell 111(6), 2002
PMID: 12526809
Aromatic amino acids at the surface of InlB are essential for host cell invasion by Listeria monocytogenes.
Machner MP, Frese S, Schubert WD, Orian-Rousseau V, Gherardi E, Wehland J, Niemann HH, Heinz DW., Mol. Microbiol. 48(6), 2003
PMID: 12791136
X-ray and neutron small-angle scattering analysis of the complex formed by the Met receptor and the Listeria monocytogenes invasion protein InlB.
Niemann HH, Petoukhov MV, Hartlein M, Moulin M, Gherardi E, Timmins P, Heinz DW, Svergun DI., J. Mol. Biol. 377(2), 2008
PMID: 18262542
Single-molecule photobleaching reveals increased MET receptor dimerization upon ligand binding in intact cells.
Dietz MS, Haße D, Ferraris DM, Gohler A, Niemann HH, Heilemann M., BMC Biophys 6(1), 2013
PMID: 23731667
Ligand-mediated dimerization of the Met receptor tyrosine kinase by the bacterial invasion protein InlB.
Ferraris DM, Gherardi E, Di Y, Heinz DW, Niemann HH., J. Mol. Biol. 395(3), 2009
PMID: 19900460
Design of a binding scaffold based on variable lymphocyte receptors of jawless vertebrates by module engineering.
Lee SC, Park K, Han J, Lee JJ, Kim HJ, Hong S, Heu W, Kim YJ, Ha JS, Lee SG, Cheong HK, Jeon YH, Kim D, Kim HS., Proc. Natl. Acad. Sci. U.S.A. 109(9), 2012
PMID: 22328160
Characterization of the calcium-binding sites of Listeria monocytogenes InlB.
Marino M, Banerjee M, Copp J, Dramsi S, Chapman T, van der Geer P, Cossart P, Ghosh P., Biochem. Biophys. Res. Commun. 316(2), 2004
PMID: 15020228
GW domains of the Listeria monocytogenes invasion protein InlB are SH3-like and mediate binding to host ligands.
Marino M, Banerjee M, Jonquieres R, Cossart P, Ghosh P., EMBO J. 21(21), 2002
PMID: 12411480
XDS.
Kabsch W., Acta Crystallogr. D Biol. Crystallogr. 66(Pt 2), 2010
PMID: 20124692
Phaser crystallographic software.
McCoy AJ, Grosse-Kunstleve RW, Adams PD, Winn MD, Storoni LC, Read RJ., J Appl Crystallogr 40(Pt 4), 2007
PMID: 19461840
Features and development of Coot.
Emsley P, Lohkamp B, Scott WG, Cowtan K., Acta Crystallogr. D Biol. Crystallogr. 66(Pt 4), 2010
PMID: 20383002
REFMAC5 for the refinement of macromolecular crystal structures.
Murshudov GN, Skubak P, Lebedev AA, Pannu NS, Steiner RA, Nicholls RA, Winn MD, Long F, Vagin AA., Acta Crystallogr. D Biol. Crystallogr. 67(Pt 4), 2011
PMID: 21460454
TLSMD web server for the generation of multi-group TLS models
AUTHOR UNKNOWN, 2006
MolProbity: all-atom structure validation for macromolecular crystallography.
Chen VB, Arendall WB 3rd, Headd JJ, Keedy DA, Immormino RM, Kapral GJ, Murray LW, Richardson JS, Richardson DC., Acta Crystallogr. D Biol. Crystallogr. 66(Pt 1), 2009
PMID: 20057044
Solution for best rotation to relate 2 sets of vectors
AUTHOR UNKNOWN, 1976
The PyMOL molecular graphics system
AUTHOR UNKNOWN, 2002
A helical RGD motif promoting cell adhesion: crystal structures of the Helicobacter pylori type IV secretion system pilus protein CagL.
Barden S, Lange S, Tegtmeyer N, Conradi J, Sewald N, Backert S, Niemann HH., Structure 21(11), 2013
PMID: 24076404
Functional map and domain structure of MET, the product of the c-met protooncogene and receptor for hepatocyte growth factor/scatter factor.
Gherardi E, Youles ME, Miguel RN, Blundell TL, Iamele L, Gough J, Bandyopadhyay A, Hartmann G, Butler PJ., Proc. Natl. Acad. Sci. U.S.A. 100(21), 2003
PMID: 14528000
Export

Markieren/ Markierung löschen
Markierte Publikationen

Open Data PUB

Web of Science

Dieser Datensatz im Web of Science®
Quellen

PMID: 24669959
PubMed | Europe PMC

Suchen in

Google Scholar