title: A structured interdomain linker directs self-polymerization of human uromodulin.
abstract: Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. Via its bipartite zona pellucida module (ZP-N/ZP-C), UMOD forms extracellular filaments that regulate kidney electrolyte balance and innate immunity, as well as protect against renal stones. Moreover, salt-dependent aggregation of UMOD filaments in the urine generates a soluble molecular net that captures uropathogenic bacteria and facilitates their clearance. Despite the functional importance of its homopolymers, no structural information is available on UMOD and how it self-assembles into filaments. Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. The structure of UMOD reveals that an extensive hydrophobic interface mediates ZP-N domain homodimerization. This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). Our data provide an example of how interdomain linker plasticity can modulate the function of structurally similar multidomain proteins. Moreover, the architecture of UMOD rationalizes numerous pathogenic mutations in both UMOD and TECTA genes.
authors.affiliation: Department of Biosciences and Nutrition & Center for Innovative Medicine, Karolinska Institutet, SE-141 83 Huddinge, Sweden;
authors.forename: Marcel
authors.initials: M
authors.lastname: Bokhove
authors.affiliation: Department of Biosciences and Nutrition & Center for Innovative Medicine, Karolinska Institutet, SE-141 83 Huddinge, Sweden;
authors.forename: Kaoru
authors.initials: K
authors.lastname: Nishimura
authors.affiliation: Molecular Genetics of Renal Disorders Unit, Division of Genetics and Cell Biology, San Raffaele Scientific Institute, I-20132 Milan, Italy;
authors.forename: Martina
authors.initials: M
authors.lastname: Brunati
authors.affiliation: Department of Biosciences and Nutrition & Center for Innovative Medicine, Karolinska Institutet, SE-141 83 Huddinge, Sweden;
authors.forename: Ling
authors.initials: L
authors.lastname: Han
authors.affiliation: European Synchrotron Radiation Facility - The European Synchrotron, Grenoble 38000, France.
authors.forename: Daniele
authors.initials: D
authors.lastname: de Sanctis
authors.affiliation: Molecular Genetics of Renal Disorders Unit, Division of Genetics and Cell Biology, San Raffaele Scientific Institute, I-20132 Milan, Italy;
authors.forename: Luca
authors.initials: L
authors.lastname: Rampoldi
authors.affiliation: Department of Biosciences and Nutrition & Center for Innovative Medicine, Karolinska Institutet, SE-141 83 Huddinge, Sweden; luca.jovine@ki.se.
authors.forename: Luca
authors.identifier: http://orcid.org/0000-0002-2679-6946
authors.initials: L
authors.lastname: Jovine
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D018360:Crystallography, X-Ray; D004220:Disulfides / Q000378:metabolism; D004285:Dogs; D016326:Extracellular Matrix Proteins / Q000235:genetics; D005455:Fluorescent Antibody Technique; D058851:GPI-Linked Proteins / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D061985:Madin Darby Canine Kidney Cells; D058619:Maltose-Binding Proteins / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020125:Mutation, Missense / Q000235:genetics; D058105:Polymerization; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016415:Sequence Alignment; D040681:Structural Homology, Protein; D058949:Uromodulin / Q000737:chemistry* / Q000648:ultrastructure
chemical_list: D004220:Disulfides; D016326:Extracellular Matrix Proteins; D058851:GPI-Linked Proteins; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins; C491350:TECTA protein, human; D058949:Uromodulin
keywords: X-ray crystallography; ZP2; polymerization; uromodulin; zona pellucida domain
references.citation: Biochemistry. 2004 Jun 15;43(23):7459-78
references.pmid: 15182189
references.citation: Nat Cell Biol. 2002 Jun;4(6):457-61
references.pmid: 12021773
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501
references.pmid: 20383002
references.citation: J Mol Biol. 2000 Sep 8;302(1):205-17
references.pmid: 10964570
references.citation: Acta Crystallogr D Biol Crystallogr. 2008 Jan;64(Pt 1):61-9
references.pmid: 18094468
references.citation: Nucleic Acids Res. 2015 Jan;43(Database issue):D257-60
references.pmid: 25300481
references.citation: J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674
references.pmid: 19461840
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21
references.pmid: 20057044
references.citation: Annu Rev Biochem. 2005;74:83-114
references.pmid: 15952882
references.citation: Cell. 2010 Oct 29;143(3):404-15
references.pmid: 20970175
references.citation: Nat Struct Mol Biol. 2015 Nov;22(11):833-4
references.pmid: 26581513
references.citation: Acta Crystallogr D Biol Crystallogr. 2012 Apr;68(Pt 4):352-67
references.pmid: 22505256
references.citation: Elife. 2015;4:e08887
references.pmid: 26673890
references.citation: Hum Genet. 2014 Jan;133(1):1-9
references.pmid: 24077912
references.citation: Mol Biol Cell. 2009 Jan;20(2):589-99
references.pmid: 19005207
references.citation: Acta Crystallogr D Biol Crystallogr. 2006 Sep;62(Pt 9):1002-11
references.pmid: 16929101
references.citation: Proc Natl Acad Sci U S A. 2004 Apr 20;101(16):5922-7
references.pmid: 15079052
references.citation: PLoS One. 2013;8(6):e67214
references.pmid: 23826237
references.citation: J Biol Chem. 2003 Sep 5;278(36):34189-202
references.pmid: 12799386
references.citation: J Cell Sci. 2011 Mar 15;124(Pt 6):940-50
references.pmid: 21378311
references.citation: Dev Growth Differ. 2002 Jun;44(3):205-12
references.pmid: 12060070
references.citation: Nat Med. 2013 Dec;19(12):1655-60
references.pmid: 24185693
references.citation: Ann Clin Lab Sci. 2010 Fall;40(4):380-5
references.pmid: 20947814
references.citation: Neuron. 2000 Oct;28(1):273-85
references.pmid: 11087000
references.citation: Science. 1985 Aug 2;229(4712):479-81
references.pmid: 2409603
references.citation: Protein Sci. 2010 May;19(5):901-13
references.pmid: 20196072
references.citation: Hum Mol Genet. 2003 Dec 15;12(24):3369-84
references.pmid: 14570709
references.citation: Protein Sci. 1998 Jul;7(7):1612-9
references.pmid: 9684894
references.citation: Nat Neurosci. 2005 Aug;8(8):1035-42
references.pmid: 15995703
references.citation: BMC Bioinformatics. 2004 Jun 4;5:69
references.pmid: 15180909
references.citation: Eur J Neurosci. 2005 Jun;21(12):3291-300
references.pmid: 16026467
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13419-24
references.pmid: 12370423
references.citation: Am J Physiol Renal Physiol. 2004 Apr;286(4):F795-802
references.pmid: 14665435
references.citation: Am J Kidney Dis. 2003 Oct;42(4):658-76
references.pmid: 14520616
references.citation: Nucleic Acids Res. 2014 Jan;42(Database issue):D222-30
references.pmid: 24288371
references.citation: Nucleic Acids Res. 2014 Jul;42(Web Server issue):W320-4
references.pmid: 24753421
references.citation: J Synchrotron Radiat. 2004 Jan 1;11(Pt 1):49-52
references.pmid: 14646132
references.citation: Am J Physiol Renal Physiol. 2010 Sep;299(3):F469-78
references.pmid: 20591941
references.citation: Proc Natl Acad Sci U S A. 2011 Mar 29;108(13):5232-6
references.pmid: 21402931
references.citation: FEBS Open Bio. 2015 May 27;5:454-65
references.pmid: 26106520
references.citation: Proc Soc Exp Biol Med. 1950 May;74(1):106-8
references.pmid: 15430405
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21
references.pmid: 20124702
references.citation: Hypertension. 2014 Mar;63(3):551-8
references.pmid: 24324041
references.citation: Nature. 2008 Dec 4;456(7222):653-7
references.pmid: 19052627
references.citation: Biochem Biophys Res Commun. 2008 Jun 6;370(3):410-3
references.pmid: 18375198
references.citation: Nat Neurosci. 2007 Feb;10(2):215-23
references.pmid: 17220887
references.citation: Mol Reprod Dev. 2008 May;75(5):847-56
references.pmid: 17926344
references.citation: Mol Biol Evol. 2011 Jan;28(1):399-406
references.pmid: 20696791
references.citation: Am J Nephrol. 2012;36(2):114-20
references.pmid: 22776760
references.citation: J Synchrotron Radiat. 2012 May;19(Pt 3):455-61
references.pmid: 22514185
references.citation: Protein Sci. 2011 Nov;20(11):1876-90
references.pmid: 21898649
references.citation: Appl Microbiol Biotechnol. 2010 Sep;88(1):187-97
references.pmid: 20535468
references.citation: Hum Mutat. 2011 May;32(5):557-63
references.pmid: 21520333
references.citation: FEBS Lett. 1992 Apr 6;300(3):237-40
references.pmid: 1313375
references.citation: Biochem J. 2004 Nov 15;384(Pt 1):191-9
references.pmid: 15264999
references.citation: PLoS One. 2009;4(6):e5936
references.pmid: 19529781
references.citation: Nephrol Dial Transplant. 2004 Dec;19(12):3150-4
references.pmid: 15575003
references.citation: J Mol Biol. 2007 Sep 21;372(3):774-97
references.pmid: 17681537
references.citation: Kidney Int. 2012 Apr;81(8):769-78
references.pmid: 22237754
references.citation: BMC Med Genet. 2014;15:34
references.pmid: 25008054
references.citation: Mol Biol Evol. 2007 Oct;24(10):2286-97
references.pmid: 17675335
references.citation: Acta Crystallogr D Biol Crystallogr. 2006 Oct;62(Pt 10):1243-50
references.pmid: 17001101
references.citation: Bioinformatics. 2000 Feb;16(2):135-9
references.pmid: 10842735
references.citation: Biochemistry. 1993 Jul 27;32(29):7334-53
references.pmid: 8338831
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):125-32
references.pmid: 20124692
references.citation: FEBS Lett. 2004 Dec 17;578(3):236-8
references.pmid: 15589826
references.citation: Methods Mol Biol. 2014;1079:263-71
references.pmid: 24170408
references.citation: Proc Natl Acad Sci U S A. 2002 Mar 5;99(5):2754-9
references.pmid: 11880627
references.citation: Kidney Int. 2011 Aug;80(4):338-47
references.pmid: 21654721
references.citation: J Immunol Methods. 1989 Jun 21;120(2):185-9
references.pmid: 2500486
references.citation: Structure. 2007 Mar;15(3):267-73
references.pmid: 17355862
references.citation: Science. 2010 Jul 9;329(5988):216-9
references.pmid: 20616279
references.citation: Biochim Biophys Acta. 2000 Apr 25;1491(1-3):376-80
references.pmid: 10760606