title: LRP1 is a receptor for Clostridium perfringens TpeL toxin indicating a two-receptor model of clostridial glycosylating toxins.
abstract: Large glycosylating toxins are major virulence factors of various species of pathogenic Clostridia. Prototypes are Clostridium difficile toxins A and B, which cause antibiotics-associated diarrhea and pseudomembranous colitis. The current model of the toxins' action suggests that receptor binding is mediated by a C-terminal domain of combined repetitive oligopeptides (CROP). This model is challenged by the glycosylating Clostridium perfringens large cytotoxin (TpeL toxin) that is devoid of the CROP domain but still intoxicates cells. Using a haploid genetic screen, we identified LDL receptor-related protein 1 (LRP1) as a host cell receptor for the TpeL toxin. LRP1-deficient cells are not able to take up TpeL and are not intoxicated. Expression of cluster IV of LRP1 is sufficient to rescue toxin uptake in these cells. By plasmon resonance spectroscopy, a KD value of 23 nM was determined for binding of TpeL to LRP1 cluster IV. The C terminus of TpeL (residues 1335-1779) represents the receptor-binding domain (RBD) of the toxin. RBD-like regions are conserved in all other clostridial glycosylating toxins preceding their CROP domain. CROP-deficient C. difficile toxin B is toxic to cells, depending on the RBD-like region (residues 1349-1811) but does not interact with LRP1. Our data indicate the presence of a second, CROP-independent receptor-binding domain in clostridial glycosylating toxins and suggest a two-receptor model for the cellular uptake of clostridial glycosylating toxins.
authors.affiliation: Institut für Experimentelle und Klinische Pharmakologie und Toxikologie, Albert-Ludwigs-Universität Freiburg, D-79104 Freiburg, Germany.
authors.forename: Björn
authors.initials: B
authors.lastname: Schorch
authors.forename: Shuo
authors.initials: S
authors.lastname: Song
authors.forename: Ferdy R
authors.initials: FR
authors.lastname: van Diemen
authors.forename: Hans H
authors.initials: HH
authors.lastname: Bock
authors.forename: Petra
authors.initials: P
authors.lastname: May
authors.forename: Joachim
authors.initials: J
authors.lastname: Herz
authors.forename: Thijn R
authors.initials: TR
authors.lastname: Brummelkamp
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Papatheodorou
authors.forename: Klaus
authors.initials: K
authors.lastname: Aktories
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000737:chemistry / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D003016:Clostridium perfringens / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000166:cytology; D004705:Endocytosis; D005347:Fibroblasts / Q000378:metabolism; D005820:Genetic Testing; D006031:Glycosylation; D006238:Haploidy; D006367:HeLa Cells; D006801:Humans; D026503:Low Density Lipoprotein Receptor-Related Protein-1 / Q000378:metabolism*; D051379:Mice; D008954:Models, Biological; D011485:Protein Binding; D017434:Protein Structure, Tertiary
chemical_list: D001427:Bacterial Toxins; D026503:Low Density Lipoprotein Receptor-Related Protein-1
keywords: receptor-mediated endocytosis; toxin receptor
references.citation: N Engl J Med. 2008 Oct 30;359(18):1932-40
references.pmid: 18971494
references.citation: J Biol Chem. 2000 Aug 18;275(33):25616-24
references.pmid: 10827173
references.citation: Proc Natl Acad Sci U S A. 1989 Aug;86(15):5810-4
references.pmid: 2762297
references.citation: FEMS Microbiol Lett. 1997 Oct 1;155(1):45-54
references.pmid: 9345763
references.citation: N Engl J Med. 2005 Dec 8;353(23):2442-9
references.pmid: 16322602
references.citation: Sci Signal. 2008 Nov 25;1(47):ra15
references.pmid: 19036715
references.citation: Nature. 1995 Jun 8;375(6531):500-3
references.pmid: 7777059
references.citation: J Biol Chem. 1999 Oct 15;274(42):30087-93
references.pmid: 10514495
references.citation: Nat Struct Mol Biol. 2006 May;13(5):460-1
references.pmid: 16622409
references.citation: Biochem Biophys Res Commun. 2003 Jan 17;300(3):706-11
references.pmid: 12507507
references.citation: Clin Microbiol Rev. 2005 Apr;18(2):247-63
references.pmid: 15831824
references.citation: Rev Physiol Biochem Pharmacol. 2004;152:23-47
references.pmid: 15449191
references.citation: J Cell Biol. 1990 Apr;110(4):1041-8
references.pmid: 1691187
references.citation: J Biol Chem. 2004 Dec 24;279(52):54944-51
references.pmid: 15489233
references.citation: Trends Microbiol. 2008 May;16(5):222-9
references.pmid: 18394902
references.citation: Gene. 1990 Nov 30;96(1):107-13
references.pmid: 2148295
references.citation: EMBO J. 2005 Aug 17;24(16):2851-61
references.pmid: 16052210
references.citation: EMBO J. 1996 Jun 3;15(11):2632-9
references.pmid: 8654360
references.citation: J Biol Chem. 1997 Sep 19;272(38):23946-51
references.pmid: 9295345
references.citation: J Biol Chem. 1992 Jun 25;267(18):12420-3
references.pmid: 1618748
references.citation: J Biol Chem. 1994 Jun 3;269(22):15827-32
references.pmid: 7515061
references.citation: J Neurosci. 2011 Feb 2;31(5):1644-51
references.pmid: 21289173
references.citation: Microb Pathog. 1994 Apr;16(4):297-303
references.pmid: 7968458
references.citation: Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1839-42
references.pmid: 8127891
references.citation: Infect Immun. 1992 Jun;60(6):2488-92
references.pmid: 1375199
references.citation: Gene Ther. 2000 Jun;7(12):1063-6
references.pmid: 10871756
references.citation: PLoS One. 2011 Mar 18;6(3):e17623
references.pmid: 21445253
references.citation: Cell. 1995 Jul 28;82(2):331-40
references.pmid: 7543026
references.citation: Nature. 2007 Mar 22;446(7134):415-9
references.pmid: 17334356
references.citation: J Biol Chem. 1996 Apr 26;271(17):10217-24
references.pmid: 8626586
references.citation: Infect Immun. 1986 Sep;53(3):573-81
references.pmid: 3744552
references.citation: Proc Natl Acad Sci U S A. 2010 Jul 27;107(30):13467-72
references.pmid: 20624955
references.citation: Proc Natl Acad Sci U S A. 2011 Sep 27;108(39):16422-7
references.pmid: 21930894
references.citation: Infect Immun. 2011 Feb;79(2):905-10
references.pmid: 21098103
references.citation: J Lipid Res. 1999 Jan;40(1):1-16
references.pmid: 9869645
references.citation: EMBO J. 1995 May 15;14(10):2269-80
references.pmid: 7774585
references.citation: Mol Microbiol. 2011 Mar;79(6):1643-54
references.pmid: 21231971
references.citation: Curr Top Microbiol Immunol. 2013;364:61-90
references.pmid: 23239349
references.citation: Infect Immun. 2008 Jul;76(7):2862-71
references.pmid: 18411291
references.citation: Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7427-31
references.pmid: 1502154
references.citation: EMBO J. 1988 Dec 20;7(13):4119-27
references.pmid: 3266596
references.citation: Biochem J. 1996 Jan 1;313 ( Pt 1):71-6
references.pmid: 8546712
references.citation: J Bacteriol. 1992 Oct;174(20):6707-10
references.pmid: 1307487
references.citation: J Clin Invest. 2001 Sep;108(6):779-84
references.pmid: 11560943
references.citation: J Biol Chem. 2007 Aug 31;282(35):25314-21
references.pmid: 17591770
references.citation: EMBO J. 1998 Aug 17;17(16):4617-25
references.pmid: 9707421
references.citation: J Biol Chem. 1991 Nov 5;266(31):21232-8
references.pmid: 1718973
references.citation: Physiol Rev. 2008 Jul;88(3):887-918
references.pmid: 18626063
references.citation: J Clin Invest. 1996 Aug 1;98(3):641-9
references.pmid: 8698855
references.citation: J Biol Chem. 1990 Oct 15;265(29):17401-4
references.pmid: 1698775
references.citation: Science. 2009 Nov 27;326(5957):1231-5
references.pmid: 19965467
references.citation: N Engl J Med. 2005 Dec 8;353(23):2433-41
references.pmid: 16322603
references.citation: PLoS One. 2010 May 17;5(5):e10673
references.pmid: 20498856
references.citation: J Biol Chem. 2012 Jul 20;287(30):24929-40
references.pmid: 22665487
references.citation: J Cell Sci. 2001 Mar;114(Pt 5):899-908
references.pmid: 11181173
references.citation: BMC Microbiol. 2008 Nov 06;8:192
references.pmid: 18990232
references.citation: Ann N Y Acad Sci. 1994 Sep 10;737:70-87
references.pmid: 7944155
references.citation: Microbiology (Reading). 2007 Apr;153(Pt 4):1198-1206
references.pmid: 17379729
references.citation: Infect Immun. 1991 Jan;59(1):73-8
references.pmid: 1670930
references.citation: Methods Cell Biol. 1994;43 Pt A:305-34
references.pmid: 7823869
references.citation: J Biol Chem. 1996 Oct 11;271(41):25173-7
references.pmid: 8810274