title: Proteomic View of Basement Membranes from Human Retinal Blood Vessels, Inner Limiting Membranes, and Lens Capsules.
abstract: Basement membranes (BMs) are extracellular matrix sheets comprising the laminins, type-IV collagens, nidogens, and the heparan sulfate proteoglycans, perlecan, collagen XVIII, and agrin. In intact BMs, BM proteins are physiologically insoluble and partially resistant to proteolytic digestion, making BMs a challenge to study. Here three types of BMs from adult human eyes, the inner limiting membrane (ILM), the retinal vascular BMs, and the lens capsule, were isolated for analysis by 1D-SDS-PAGE and LC-MS/MS. Peptide and protein identifications were done using MaxQuant. 1129 proteins were identified with a 1% false discovery rate. Data showed that BMs are composed of multiple laminins, collagen IVs, nidogens, and proteoglycans. The dominant laminin family member in all BMs was laminin α5β2γ1. The dominant collagen IV trimer in lens capsule (LC) and blood vessel (BV) BMs had a chain composition of α1(IV)2, α2 (IV), whereas the dominant collagen IV in the ILM had the α3(IV), α4(IV), α5(IV) chain composition. The data also showed that the ratio of laminin and collagen IVs varied among different BM types: the ratio of collagen IV to the other BM proteins is highest in LC, followed by BV and lowest for the ILM. The data have been deposited to the ProteomeXchange with identifier PXD001025.
authors.affiliation: Proteomics Core, Genomics and Proteomics Core laboratories, University of Pittsburgh , 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15260, United States.
authors.forename: Guy
authors.initials: G
authors.lastname: Uechi
authors.affiliation: Proteomics Core, Genomics and Proteomics Core laboratories, University of Pittsburgh , 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15260, United States.
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Sun
authors.affiliation: Proteomics Core, Genomics and Proteomics Core laboratories, University of Pittsburgh , 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15260, United States.
authors.forename: Emanuel M
authors.initials: EM
authors.lastname: Schreiber
authors.affiliation: Department of Neurobiology, University of Pittsburgh , 200 Lothrop Street, Pittsburgh, Pennsylvania 15261, United States.
authors.forename: Willi
authors.initials: W
authors.lastname: Halfter
authors.affiliation: Proteomics Core, Genomics and Proteomics Core laboratories, University of Pittsburgh , 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15260, United States.
authors.forename: Manimalha
authors.initials: M
authors.lastname: Balasubramani
keywords: basement membrane; collagen IV; collagens; laminin; mass spectrometry; nidogens; perlecan; proteomics