6dbf

From Proteopedia

(Difference between revisions)

Current revision

Crystal Structure of VHH R303 in complex with InlB-LRR

Structural highlights

6dbf is a 2 chain structure with sequence from Camelus dromedarius and Listeria monocytogenes. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:	X-ray diffraction, Resolution 1.55Å
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

INLB_LISMG Mediates the entry of L.monocytogenes into normally non-phagocytic mammalian host cells (Probable) (PubMed:11081636, PubMed:9282740). Its host receptor is hepatocyte growth factor receptor (HGF receptor, a tyrosine kinase, MET) which is tyrosine-phosphorylated in response to InlB in human, green monkey, mouse and dog cell lines (PubMed:11081636, PubMed:15049825). Downstream adapter proteins GAB1 and CBL are phosphorylated in response to InlB, which also causes cell colony scattering (PubMed:11081636). InlB binding to mammalian cells is saturable and inhibited by EDTA; InlB-coated beads can be taken up by host cells (PubMed:10747014). Complement component 1 Q subcomponent-binding protein (gC1q-R, C1QBP) might act as an InlB receptor, leading to activation of PI3-kinase in green monkey cells (PubMed:10747014). Stimulation of Tyr-phosphorylation by InlB is antagonized by C1QBP, showing that potentiation of MET signaling via the GW domains is not mediated by C1QBP; the exact role of C1QBP remains to be determined (PubMed:15049825). Stimulation of Tyr-phosphorylation of MET by InlB is potentiated by the InlB GW domains and glycosaminoglycans such as heparin; exogenously added InlB, or hepatocyte growth factor (HGF) will also substitute for bacterial InlB, suggesting InlB promotes bacterial invasion by mimicking the hormone HGF (PubMed:15049825). May stimulate phosphatidylinositol 4,5-bisphosphate 3-kinase (PI3-kinase) in green monkey cells, has less effect in humans as PI3-kinase is constitutively and highly expressed in Caco cells (Probable). Binds heparin; C1QBP and heparin seem to bind to the GW domains (PubMed:12411480).^[1] ^[2] ^[3] ^[4] ^[5] ^[6] ^[7] ^[8]

Publication Abstract from PubMed

Listeria monocytogenes causes Listeriosis, a potentially fatal food borne disease. The condition is especially harmful to pregnant women. Listeria outbreaks can originate from diverse foods, highlighting the need for novel strategies to improve food safety. The first step in Listeria invasion is internalization of the bacteria, which is mediated by the interaction of the internalin family of virulence factors with host cell receptors. A crucial interaction for Listeria invasion of the placenta, and thus is a target for therapeutic intervention, is between Internalin B (InlB) and the receptor c-Met.. Single-domain antibodies (VHH, also called nanobodies, or sdAbs) from camel heavy-chain antibodies are a novel solution for preventing Listeria infections. The VHH R303, R330 and R326 all bind InlB with high affinity, however the molecular mechanism behind their mode of action was unknown. We demonstrate that despite a high degree of sequence and structural diversity, the VHH bind a single epitope on InlB. A combination of gentamicin protection assays and florescent microscopy establish that InlB specific VHH inhibit Listeria invasion of HeLa cells. A high resolution X-ray structure of VHH R303 in complex with InlB showed that the VHH binds at the c-Met interaction site on InlB, thereby acting as a competitive inhibitor preventing bacterial invasion. These results point to the potential of VHH as a novel class of therapeutics for the prevention of Listeriosis.

Structural Basis of VHH mediated neutralization of the foodborne pathogen Listeria monocytogenes.,Toride King M, Huh I, Shenai A, Brooks TM, Brooks CL J Biol Chem. 2018 Jul 5. pii: RA118.003888. doi: 10.1074/jbc.RA118.003888. PMID:29976754^[9]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Braun L, Ghebrehiwet B, Cossart P. gC1q-R/p32, a C1q-binding protein, is a receptor for the InlB invasion protein of Listeria monocytogenes. EMBO J. 2000 Apr 3;19(7):1458-66. PMID:10747014 doi:10.1093/emboj/19.7.1458
↑ Shen Y, Naujokas M, Park M, Ireton K. InIB-dependent internalization of Listeria is mediated by the Met receptor tyrosine kinase. Cell. 2000 Oct 27;103(3):501-10. PMID:11081636 doi:10.1016/s0092-8674(00)00141-0
↑ Marino M, Banerjee M, Jonquieres R, Cossart P, Ghosh P. GW domains of the Listeria monocytogenes invasion protein InlB are SH3-like and mediate binding to host ligands. EMBO J. 2002 Nov 1;21(21):5623-34. PMID:12411480
↑ Banerjee M, Copp J, Vuga D, Marino M, Chapman T, van der Geer P, Ghosh P. GW domains of the Listeria monocytogenes invasion protein InlB are required for potentiation of Met activation. Mol Microbiol. 2004 Apr;52(1):257-71. PMID:15049825 doi:10.1111/j.1365-2958.2003.03968.x
↑ Braun L, Dramsi S, Dehoux P, Bierne H, Lindahl G, Cossart P. InlB: an invasion protein of Listeria monocytogenes with a novel type of surface association. Mol Microbiol. 1997 Jul;25(2):285-94. PMID:9282740 doi:10.1046/j.1365-2958.1997.4621825.x
↑ Shen Y, Naujokas M, Park M, Ireton K. InIB-dependent internalization of Listeria is mediated by the Met receptor tyrosine kinase. Cell. 2000 Oct 27;103(3):501-10. PMID:11081636 doi:10.1016/s0092-8674(00)00141-0
↑ Gaillard JL, Berche P, Frehel C, Gouin E, Cossart P. Entry of L. monocytogenes into cells is mediated by internalin, a repeat protein reminiscent of surface antigens from gram-positive cocci. Cell. 1991 Jun 28;65(7):1127-41. PMID:1905979 doi:10.1016/0092-8674(91)90009-n
↑ Ireton K, Payrastre B, Chap H, Ogawa W, Sakaue H, Kasuga M, Cossart P. A role for phosphoinositide 3-kinase in bacterial invasion. Science. 1996 Nov 1;274(5288):780-2. PMID:8864117 doi:10.1126/science.274.5288.780
↑ Toride King M, Huh I, Shenai A, Brooks TM, Brooks CL. Structural Basis of VHH mediated neutralization of the foodborne pathogen Listeria monocytogenes. J Biol Chem. 2018 Jul 5. pii: RA118.003888. doi: 10.1074/jbc.RA118.003888. PMID:29976754 doi:http://dx.doi.org/10.1074/jbc.RA118.003888

1 Structural highlights
2 Function
3 Publication Abstract from PubMed
4 References

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/6dbf"

@@ Line 3: / Line 3: @@
 <StructureSection load='6dbf' size='340' side='right'caption='[[6dbf]], [[Resolution|resolution]] 1.55&Aring;' scene=''>
 == Structural highlights ==
-<table><tr><td colspan='2'>[[6dbf]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/"bacterium_monocytogenes_hominis"_nyfeldt_1932 "bacterium monocytogenes hominis" nyfeldt 1932] and [http://en.wikipedia.org/wiki/Arabian_camel Arabian camel]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6DBF OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6DBF FirstGlance]. <br>
+<table><tr><td colspan='2'>[[6dbf]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Camelus_dromedarius Camelus dromedarius] and [https://en.wikipedia.org/wiki/Listeria_monocytogenes Listeria monocytogenes]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6DBF OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6DBF FirstGlance]. <br>
-</td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[6dba|6dba]], [[6dbd|6dbd]], [[6dbe|6dbe]]</td></tr>
+</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.55&#8491;</td></tr>
-<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6dbf FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6dbf OCA], [http://pdbe.org/6dbf PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6dbf RCSB], [http://www.ebi.ac.uk/pdbsum/6dbf PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6dbf ProSAT]</span></td></tr>
+<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=6dbf FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6dbf OCA], [https://pdbe.org/6dbf PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6dbf RCSB], [https://www.ebi.ac.uk/pdbsum/6dbf PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6dbf ProSAT]</span></td></tr>
 </table>
+== Function ==
+[https://www.uniprot.org/uniprot/INLB_LISMG INLB_LISMG] Mediates the entry of L.monocytogenes into normally non-phagocytic mammalian host cells (Probable) (PubMed:11081636, PubMed:9282740). Its host receptor is hepatocyte growth factor receptor (HGF receptor, a tyrosine kinase, MET) which is tyrosine-phosphorylated in response to InlB in human, green monkey, mouse and dog cell lines (PubMed:11081636, PubMed:15049825). Downstream adapter proteins GAB1 and CBL are phosphorylated in response to InlB, which also causes cell colony scattering (PubMed:11081636). InlB binding to mammalian cells is saturable and inhibited by EDTA; InlB-coated beads can be taken up by host cells (PubMed:10747014). Complement component 1 Q subcomponent-binding protein (gC1q-R, C1QBP) might act as an InlB receptor, leading to activation of PI3-kinase in green monkey cells (PubMed:10747014). Stimulation of Tyr-phosphorylation by InlB is antagonized by C1QBP, showing that potentiation of MET signaling via the GW domains is not mediated by C1QBP; the exact role of C1QBP remains to be determined (PubMed:15049825). Stimulation of Tyr-phosphorylation of MET by InlB is potentiated by the InlB GW domains and glycosaminoglycans such as heparin; exogenously added InlB, or hepatocyte growth factor (HGF) will also substitute for bacterial InlB, suggesting InlB promotes bacterial invasion by mimicking the hormone HGF (PubMed:15049825). May stimulate phosphatidylinositol 4,5-bisphosphate 3-kinase (PI3-kinase) in green monkey cells, has less effect in humans as PI3-kinase is constitutively and highly expressed in Caco cells (Probable). Binds heparin; C1QBP and heparin seem to bind to the GW domains (PubMed:12411480).<ref>PMID:10747014</ref> <ref>PMID:11081636</ref> <ref>PMID:12411480</ref> <ref>PMID:15049825</ref> <ref>PMID:9282740</ref> <ref>PMID:11081636</ref> <ref>PMID:1905979</ref> <ref>PMID:8864117</ref>
 <div style="background-color:#fffaf0;">
 == Publication Abstract from PubMed ==
@@ Line 20: / Line 22: @@
 __TOC__
 </StructureSection>
-[[Category: Bacterium monocytogenes hominis nyfeldt 1932]]
+[[Category: Camelus dromedarius]]
-[[Category: Arabian camel]]
 [[Category: Large Structures]]
-[[Category: Brooks, C L]]
+[[Category: Listeria monocytogenes]]
-[[Category: Huh, I]]
+[[Category: Brooks CL]]
-[[Category: King, M Toride]]
+[[Category: Huh I]]
-[[Category: Immune system]]
+[[Category: Toride King M]]
-[[Category: Nanobody vhh listeria internalin]]

6dbf

From Proteopedia

Current revision

Crystal Structure of VHH R303 in complex with InlB-LRR

Structural highlights

Function

Publication Abstract from PubMed

References

Contents

Proteopedia Page Contributors and Editors (what is this?)

Views

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