title: Global analysis reveals the complexity of the human glomerular extracellular matrix.
abstract: The glomerulus contains unique cellular and extracellular matrix (ECM) components, which are required for intact barrier function. Studies of the cellular components have helped to build understanding of glomerular disease; however, the full composition and regulation of glomerular ECM remains poorly understood. We used mass spectrometry-based proteomics of enriched ECM extracts for a global analysis of human glomerular ECM in vivo and identified a tissue-specific proteome of 144 structural and regulatory ECM proteins. This catalog includes all previously identified glomerular components plus many new and abundant components. Relative protein quantification showed a dominance of collagen IV, collagen I, and laminin isoforms in the glomerular ECM together with abundant collagen VI and TINAGL1. Protein network analysis enabled the creation of a glomerular ECM interactome, which revealed a core of highly connected structural components. More than one half of the glomerular ECM proteome was validated using colocalization studies and data from the Human Protein Atlas. This study yields the greatest number of ECM proteins relative to previous investigations of whole glomerular extracts, highlighting the importance of sample enrichment. It also shows that the composition of glomerular ECM is far more complex than previously appreciated and suggests that many more ECM components may contribute to glomerular development and disease processes. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium with the dataset identifier PXD000456.
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research and Faculty of Medical and Human Sciences, University of Manchester, Manchester, United Kingdom; Rachel.Lennon@manchester.ac.uk.
authors.forename: Rachel
authors.initials: R
authors.lastname: Lennon
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research and.
authors.forename: Adam
authors.initials: A
authors.lastname: Byron
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research and.
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Humphries
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research and Faculty of Medical and Human Sciences, University of Manchester, Manchester, United Kingdom;
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Randles
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research and.
authors.forename: Alex
authors.initials: A
authors.lastname: Carisey
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research and Faculty of Medical and Human Sciences, University of Manchester, Manchester, United Kingdom;
authors.forename: Stephanie
authors.initials: S
authors.lastname: Murphy
authors.affiliation: Biological Mass Spectrometry Core Facility, Faculty of Life Sciences, and.
authors.forename: David
authors.initials: D
authors.lastname: Knight
authors.affiliation: Faculty of Medical and Human Sciences, University of Manchester, Manchester, United Kingdom;
authors.forename: Paul E
authors.initials: PE
authors.lastname: Brenchley
authors.affiliation: Division of Nephrology, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee; and Department of Medicine, Veterans Affairs Hospital, Nashville, Tennessee.
authors.forename: Roy
authors.initials: R
authors.lastname: Zent
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research and.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Humphries
mesh_terms: D000328:Adult; D024142:Collagen Type VI / Q000737:chemistry; D016326:Extracellular Matrix Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D063990:Gene Ontology; D006801:Humans; D007678:Kidney Glomerulus / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D054834:Lipocalins / Q000737:chemistry; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D060066:Protein Interaction Maps; D020543:Proteome / Q000737:chemistry* / Q000235:genetics
chemical_list: D024142:Collagen Type VI; D016326:Extracellular Matrix Proteins; D054834:Lipocalins; D020543:Proteome; C423605:TINAGL1 protein, human