title: Lysyl Oxidase-like-2 Cross-links Collagen IV of Glomerular Basement Membrane.
abstract: The 7S dodecamer is recognized as an important structural cross-linking domain of collagen IV networks that provide mechanical stability to basement membranes, a specialized form of extracellular matrix essential for the development and maintenance of tissue architecture. Although the 7S dodecamer is stabilized by covalent cross-linking, the molecular mechanism by which such cross-links are formed has not been revealed. Here, we aimed to identify the enzyme(s) that cross-links the 7S dodecamer and characterize its expression in the kidney glomerulus. Pharmacological inhibition of candidate extracellular matrix enzymes revealed that lysyl oxidase activity is required for cross-linking of 7S polypeptides. Among all lysyl oxidase family members, lysyl oxidase-like-2 (LOXL2) was identified as the isoform cross-linking collagen IV in mouse embryonal PFHR-9 cells. Biochemical analyses revealed that LOXL2 readily promoted the formation of lysyl-derived cross-links in the 7S dodecamer but not in the NC1 domain. We also established that LOXL2 is the main lysyl oxidase family member present in the glomerular extracellular matrix. Altogether, we demonstrate that LOXL2 is a novel component of the molecular machinery that forms cross-linked collagen IV networks, which are essential for glomerular basement membrane stability and molecular ultrafiltration function.
authors: From the Division of Nephrology and Hypertension, Department of Medicine, and.|Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, Tennessee 37232.
authors: Carolina
authors: C
authors: Añazco
authors: From the Division of Nephrology and Hypertension, Department of Medicine, and.|Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, Tennessee 37232.
authors: Alberto J
authors: AJ
authors: López-Jiménez
authors: From the Division of Nephrology and Hypertension, Department of Medicine, and.
authors: Mohamed
authors: M
authors: Rafi
authors: From the Division of Nephrology and Hypertension, Department of Medicine, and.|Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, Tennessee 37232.
authors: Lorenzo
authors: L
authors: Vega-Montoto
authors: From the Division of Nephrology and Hypertension, Department of Medicine, and.
authors: Ming-Zhi
authors: MZ
authors: Zhang
authors: From the Division of Nephrology and Hypertension, Department of Medicine, and.|Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, Tennessee 37232.
authors: Billy G
authors: BG
authors: Hudson
authors: From the Division of Nephrology and Hypertension, Department of Medicine, and roberto.vanacore@vanderbilt.edu.|Center for Matrix Biology, Vanderbilt University Medical Center, Nashville, Tennessee 37232.
authors: Roberto M
authors: RM
authors: Vanacore
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D024141:Collagen Type IV / Q000235:genetics / Q000378:metabolism*; D005109:Extracellular Matrix / Q000235:genetics / Q000378:metabolism*; D050533:Glomerular Basement Membrane / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D051379:Mice
chemical_list: D024141:Collagen Type IV; D000594:Amino Acid Oxidoreductases; C119091:LOXL2 protein, human; C485601:Loxl2 protein, mouse
keywords: basement membrane; collagen; extracellular matrix; kidney; lysyl oxidase; protein cross-linking