title: Extracellular chloride signals collagen IV network assembly during basement membrane formation.
abstract: Basement membranes are defining features of the cellular microenvironment; however, little is known regarding their assembly outside cells. We report that extracellular Cl(-) ions signal the assembly of collagen IV networks outside cells by triggering a conformational switch within collagen IV noncollagenous 1 (NC1) domains. Depletion of Cl(-) in cell culture perturbed collagen IV networks, disrupted matrix architecture, and repositioned basement membrane proteins. Phylogenetic evidence indicates this conformational switch is a fundamental mechanism of collagen IV network assembly throughout Metazoa. Using recombinant triple helical protomers, we prove that NC1 domains direct both protomer and network assembly and show in Drosophila that NC1 architecture is critical for incorporation into basement membranes. These discoveries provide an atomic-level understanding of the dynamic interactions between extracellular Cl(-) and collagen IV assembly outside cells, a critical step in the assembly and organization of basement membranes that enable tissue architecture and function. Moreover, this provides a mechanistic framework for understanding the molecular pathobiology of NC1 domains.
authors.affiliation: Department of Biochemistry, Vanderbilt University Medical Center, Nashville, TN 37232 Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Christopher F
authors.initials: CF
authors.lastname: Cummings
authors.affiliation: Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Vadim
authors.initials: V
authors.lastname: Pedchenko
authors.affiliation: Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Structural Biology, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Kyle L
authors.initials: KL
authors.lastname: Brown
authors.affiliation: Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, TN 37232 Aspirnaut Program, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Selene
authors.initials: S
authors.lastname: Colon
authors.affiliation: Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Rafi
authors.affiliation: Aspirnaut Program, Vanderbilt University Medical Center, Nashville, TN 37232 Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Celestial
authors.initials: C
authors.lastname: Jones-Paris
authors.affiliation: Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Elena
authors.initials: E
authors.lastname: Pokydeshava
authors.affiliation: School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Min
authors.initials: M
authors.lastname: Liu
authors.affiliation: School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Jose C
authors.initials: JC
authors.lastname: Pastor-Pareja
authors.affiliation: Department of Biology, Vanderbilt University Medical Center, Nashville, TN 37232 Aspirnaut Program, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Cody
authors.initials: C
authors.lastname: Stothers
authors.affiliation: Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, TN 37232 Aspirnaut Program, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Isi A
authors.initials: IA
authors.lastname: Ero-Tolliver
authors.affiliation: Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: A Scott
authors.initials: AS
authors.lastname: McCall
authors.affiliation: Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Roberto
authors.initials: R
authors.lastname: Vanacore
authors.affiliation: Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Gautam
authors.initials: G
authors.lastname: Bhave
authors.affiliation: Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Samuel
authors.initials: S
authors.lastname: Santoro
authors.affiliation: Department of Medicine, Division of Allergy, Pulmonary, and Critical Care Medicine, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Blackwell
authors.affiliation: Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, TN 37232 Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232 Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Roy
authors.initials: R
authors.lastname: Zent
authors.affiliation: Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, TN 37232 Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, TN 37232 Department of Molecular Physiology and Biophysics, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Ambra
authors.initials: A
authors.lastname: Pozzi
authors.affiliation: Department of Biochemistry, Vanderbilt University Medical Center, Nashville, TN 37232 Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN 37232 Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, TN 37232 Aspirnaut Program, Vanderbilt University Medical Center, Nashville, TN 37232 Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232 Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232 Vanderbilt Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, TN 37232 Vanderbilt Institute of Chemical Biology, Vanderbilt University Medical Center, Nashville, TN 37232 billy.hudson@vanderbilt.edu.
authors.forename: Billy G
authors.initials: BG
authors.lastname: Hudson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001485:Basement Membrane / Q000378:metabolism* / Q000502:physiology*; D002417:Cattle; D045744:Cell Line, Tumor; D002712:Chlorides / Q000378:metabolism*; D024141:Collagen Type IV / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D010802:Phylogeny; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000235:genetics
chemical_list: D002712:Chlorides; D024141:Collagen Type IV; D021122:Protein Subunits