title: Structural decoding of netrin-4 reveals a regulatory function towards mature basement membranes.
abstract: Netrins, a family of laminin-related molecules, have been proposed to act as guidance cues either during nervous system development or the establishment of the vascular system. This was clearly demonstrated for netrin-1 via its interaction with the receptors DCC and UNC5s. However, mainly based on shared homologies with netrin-1, netrin-4 was also proposed to play a role in neuronal outgrowth and developmental/pathological angiogenesis via interactions with netrin-1 receptors. Here, we present the high-resolution structure of netrin-4, which shows unique features in comparison with netrin-1, and show that it does not bind directly to any of the known netrin-1 receptors. We show that netrin-4 disrupts laminin networks and basement membranes (BMs) through high-affinity binding to the laminin γ1 chain. We hypothesize that this laminin-related function is essential for the previously described effects on axon growth promotion and angiogenesis. Our study unveils netrin-4 as a non-enzymatic extracellular matrix protein actively disrupting pre-existing BMs.
authors.affiliation: Institute for Dental Research and Oral Musculoskeletal Biology, Medical Faculty, University of Cologne, Joseph-Stelzmann-Strasse 52, Cologne 50931, Germany.|Center for Biochemistry, Medical Faculty, University of Cologne, Cologne, Joseph-Stelzmann-Strasse 52, Cologne 50931, Germany.
authors.forename: Raphael
authors.identifier: 0000-0002-9434-4108
authors.initials: R
authors.lastname: Reuten
authors.affiliation: Alberta RNA Research and Training Institute, Department of Chemistry and Biochemistry, University of Lethbridge, 4401 University Drive, Lethbridge, Alberta T1K 3M4, Canada.|School of Biosciences, University of Birmingham, Edgbaston B15 2TT, UK.
authors.forename: Trushar R
authors.identifier: 0000-0003-0627-2923
authors.initials: TR
authors.lastname: Patel
authors.affiliation: Department of Chemistry, University of Manitoba, Winnipeg R3T 2N2, Canada.
authors.forename: Matthew
authors.initials: M
authors.lastname: McDougall
authors.affiliation: Apoptosis, Cancer and Development Laboratory, Equipe labellisée 'La Ligue', LabEx DEVweCAN, Centre de Recherche en Cancérologie de Lyon, INSERM U1052-CNRS UMR5286, Université de Lyon, Centre Léon Bérard, Lyon 69008, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Rama
authors.affiliation: Institute for Dental Research and Oral Musculoskeletal Biology, Medical Faculty, University of Cologne, Joseph-Stelzmann-Strasse 52, Cologne 50931, Germany.|Center for Biochemistry, Medical Faculty, University of Cologne, Cologne, Joseph-Stelzmann-Strasse 52, Cologne 50931, Germany.
authors.forename: Denise
authors.initials: D
authors.lastname: Nikodemus
authors.affiliation: Apoptosis, Cancer and Development Laboratory, Equipe labellisée 'La Ligue', LabEx DEVweCAN, Centre de Recherche en Cancérologie de Lyon, INSERM U1052-CNRS UMR5286, Université de Lyon, Centre Léon Bérard, Lyon 69008, France.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Gibert
authors.affiliation: Apoptosis, Cancer and Development Laboratory, Equipe labellisée 'La Ligue', LabEx DEVweCAN, Centre de Recherche en Cancérologie de Lyon, INSERM U1052-CNRS UMR5286, Université de Lyon, Centre Léon Bérard, Lyon 69008, France.
authors.forename: Jean-Guy
authors.initials: JG
authors.lastname: Delcros
authors.affiliation: Center for Applied Tissue Engineering and Regenerative Medicine-CANTER, Munich University of Applied Sciences, Lothstrasse 34, Munich D-80335, Germany.|Laboratory of Experimental Surgery and Regenerative Medicine - ExperiMed, Department of Surgery, Clinical Center University of Munich, Nussbaumstrasse 20, Munich D-80336, Germany.|Center for Nanoscience-CeNS, Geschwister-Scholl-Platz 1, Munich D-80539, Germany.
authors.forename: Carina
authors.initials: C
authors.lastname: Prein
authors.affiliation: Department of Chemistry, University of Manitoba, Winnipeg R3T 2N2, Canada.
authors.forename: Markus
authors.identifier: 0000-0003-1068-746X
authors.initials: M
authors.lastname: Meier
authors.affiliation: Laboratory for Cell and Tissue Engineering, Department of Obstetrics, University Hospital Zurich, Schmelzbergstr. 12, Zurich 8091, Switzerland.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Metzger
authors.affiliation: School of Biological Sciences, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ, UK.|Norwich Medical School, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ, UK.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: Institute for Dental Research and Oral Musculoskeletal Biology, Medical Faculty, University of Cologne, Joseph-Stelzmann-Strasse 52, Cologne 50931, Germany.|Center for Biochemistry, Medical Faculty, University of Cologne, Cologne, Joseph-Stelzmann-Strasse 52, Cologne 50931, Germany.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Kaltenberg
authors.affiliation: Department of Pathology, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.
authors.forename: Karen K
authors.initials: KK
authors.lastname: McKee
authors.affiliation: Department of Dermatology, University Hospital Magdeburg, Magdeburg 39120, Germany.|Laboratory of Experimental Dermatology, Department of Dermatology and Allergy, University of Bonn, Bonn 53105, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Bald
authors.affiliation: Department of Dermatology, University Hospital Magdeburg, Magdeburg 39120, Germany.|Laboratory of Experimental Dermatology, Department of Dermatology and Allergy, University of Bonn, Bonn 53105, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Tüting
authors.affiliation: Department of Dermatology and Venerology, University of Cologne, Cologne 50931, Germany.
authors.forename: Paola
authors.initials: P
authors.lastname: Zigrino
authors.affiliation: Institute of Anatomy, University of Bern, Baltzerstrasse 2, Bern 3000, Switzerland.
authors.forename: Valentin
authors.initials: V
authors.lastname: Djonov
authors.affiliation: Institute of Cardiovascular Research and Sport Medicine, Department of Molecular and Cellular Sport Medicine, German Sport University Cologne, Cologne 50933, Germany.
authors.forename: Wilhelm
authors.initials: W
authors.lastname: Bloch
authors.affiliation: Center for Applied Tissue Engineering and Regenerative Medicine-CANTER, Munich University of Applied Sciences, Lothstrasse 34, Munich D-80335, Germany.|Center for Nanoscience-CeNS, Geschwister-Scholl-Platz 1, Munich D-80539, Germany.
authors.forename: Hauke
authors.initials: H
authors.lastname: Clausen-Schaumann
authors.affiliation: School of Biological Sciences, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ, UK.
authors.forename: Ernst
authors.initials: E
authors.lastname: Poschl
authors.affiliation: Department of Pathology, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.
authors.forename: Peter D
authors.initials: PD
authors.lastname: Yurchenco
authors.affiliation: Laboratory for Cell and Tissue Engineering, Department of Obstetrics, University Hospital Zurich, Schmelzbergstr. 12, Zurich 8091, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Ehrbar
authors.affiliation: Apoptosis, Cancer and Development Laboratory, Equipe labellisée 'La Ligue', LabEx DEVweCAN, Centre de Recherche en Cancérologie de Lyon, INSERM U1052-CNRS UMR5286, Université de Lyon, Centre Léon Bérard, Lyon 69008, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Mehlen
authors.affiliation: Department of Chemistry, University of Manitoba, Winnipeg R3T 2N2, Canada.
authors.forename: Jörg
authors.initials: J
authors.lastname: Stetefeld
authors.affiliation: Institute for Dental Research and Oral Musculoskeletal Biology, Medical Faculty, University of Cologne, Joseph-Stelzmann-Strasse 52, Cologne 50931, Germany.|Center for Biochemistry, Medical Faculty, University of Cologne, Cologne, Joseph-Stelzmann-Strasse 52, Cologne 50931, Germany.
authors.forename: Manuel
authors.identifier: 0000-0002-2962-7814
authors.initials: M
authors.lastname: Koch
mesh_terms: D000818:Animals; D000071437:Axon Guidance / Q000502:physiology*; D001369:Axons / Q000502:physiology; D001485:Basement Membrane / Q000378:metabolism*; D002645:Chickens; D049033:Chorioallantoic Membrane / Q000502:physiology; D018360:Crystallography, X-Ray; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D007797:Laminin / Q000502:physiology*; D008545:Melanoma / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D016297:Mutagenesis, Site-Directed; D018919:Neovascularization, Physiologic / Q000502:physiology*; D000075385:Netrins / Q000502:physiology* / Q000648:ultrastructure; D011485:Protein Binding; D055503:Protein Multimerization; D051381:Rats; D017207:Rats, Sprague-Dawley; D012583:Schwann Cells; D023041:Xenograft Model Antitumor Assays
chemical_list: D007797:Laminin; D000075385:Netrins; C416214:Ntn4 protein, mouse; C089141:laminin gamma 1