title: A unique covalent bond in basement membrane is a primordial innovation for tissue evolution.
abstract: Basement membrane, a specialized ECM that underlies polarized epithelium of eumetazoans, provides signaling cues that regulate cell behavior and function in tissue genesis and homeostasis. A collagen IV scaffold, a major component, is essential for tissues and dysfunctional in several diseases. Studies of bovine and Drosophila tissues reveal that the scaffold is stabilized by sulfilimine chemical bonds (S = N) that covalently cross-link methionine and hydroxylysine residues at the interface of adjoining triple helical protomers. Peroxidasin, a heme peroxidase embedded in the basement membrane, produces hypohalous acid intermediates that oxidize methionine, forming the sulfilimine cross-link. We explored whether the sulfilimine cross-link is a fundamental requirement in the genesis and evolution of epithelial tissues by determining its occurrence and evolutionary origin in Eumetazoa and its essentiality in zebrafish development; 31 species, spanning 11 major phyla, were investigated for the occurrence of the sulfilimine cross-link by electrophoresis, MS, and multiple sequence alignment of de novo transcriptome and available genomic data for collagen IV and peroxidasin. The results show that the cross-link is conserved throughout Eumetazoa and arose at the divergence of Porifera and Cnidaria over 500 Mya. Also, peroxidasin, the enzyme that forms the bond, is evolutionarily conserved throughout Metazoa. Morpholino knockdown of peroxidasin in zebrafish revealed that the cross-link is essential for organogenesis. Collectively, our findings establish that the triad-a collagen IV scaffold with sulfilimine cross-links, peroxidasin, and hypohalous acids-is a primordial innovation of the ECM essential for organogenesis and tissue evolution.
authors.affiliation: Department of Medicine, Division of Nephrology and Hypertension, Center for Matrix Biology, Department of Biochemistry, Mass Spectrometry Research Center, Vanderbilt-Ingram Cancer Center, Vanderbilt Technologies for Advanced Genomics, Department of Medical Education and Administration, Department of Pathology, Microbiology, and Immunology, and Vanderbilt Institute of Chemical Biology, Vanderbilt University Medical Center, Nashville, TN 37232.
authors.forename: Aaron L
authors.initials: AL
authors.lastname: Fidler
authors.forename: Roberto M
authors.initials: RM
authors.lastname: Vanacore
authors.forename: Sergei V
authors.initials: SV
authors.lastname: Chetyrkin
authors.forename: Vadim K
authors.initials: VK
authors.lastname: Pedchenko
authors.forename: Gautam
authors.initials: G
authors.lastname: Bhave
authors.forename: Viravuth P
authors.initials: VP
authors.lastname: Yin
authors.forename: Cody L
authors.initials: CL
authors.lastname: Stothers
authors.forename: Kristie Lindsey
authors.initials: KL
authors.lastname: Rose
authors.forename: W Hayes
authors.initials: WH
authors.lastname: McDonald
authors.forename: Travis A
authors.initials: TA
authors.lastname: Clark
authors.forename: Dorin-Bogdan
authors.initials: DB
authors.lastname: Borza
authors.forename: Robert E
authors.initials: RE
authors.lastname: Steele
authors.forename: Michael T
authors.initials: MT
authors.lastname: Ivy
authors.forename: Julie K
authors.initials: JK
authors.lastname: Hudson
authors.forename: Billy G
authors.initials: BG
authors.lastname: Hudson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001485:Basement Membrane / Q000378:metabolism*; D005075:Biological Evolution; D024141:Collagen Type IV / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry; D004331:Drosophila melanogaster; D005109:Extracellular Matrix / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000737:chemistry; D006418:Heme / Q000737:chemistry; D007097:Imines / Q000737:chemistry*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D009195:Peroxidase / Q000737:chemistry; D010544:Peroxidases / Q000737:chemistry; D017434:Protein Structure, Tertiary; D017423:Sequence Analysis, RNA; D017386:Sequence Homology, Amino Acid; D013457:Sulfur Compounds / Q000737:chemistry*; D015027:Zebrafish; D000097423:Peroxidasin
chemical_list: D024141:Collagen Type IV; D003432:Cross-Linking Reagents; D016326:Extracellular Matrix Proteins; D006418:Heme; D007097:Imines; D010455:Peptides; D009195:Peroxidase; D010544:Peroxidases; D013457:Sulfur Compounds; D000097423:Peroxidasin; C586946:sulfilimine