title: A proteomic approach for identification and localization of the pericellular components of chondrocytes.
abstract: Although the pericellular matrix (PCM) plays a central role in the communication between chondrocytes and extracellular matrix, its composition is largely unknown. In this study, the PCM was investigated with a proteomic approach using chondrons, which are enzymatically isolated constructs including the chondrocyte and its surrounding PCM. Chondrons and chondrocytes alone were isolated from human articular cartilage. Proteins extracted from chondrons and chondrocytes were used for two-dimensional electrophoresis. Protein spots were quantitatively compared between chondron and chondrocyte gels. Cellular proteins, which had similar density between chondron and chondrocyte gels, did not proceed for analysis. Since chondrons only differ from chondrocytes in association of the PCM, protein spots in the chondron gels that had higher quantity than that in the chondrocyte gels were selected as candidates of the PCM components and processed for mass spectrometry. Among 15 identified peptides, several were fragments of the three type VI collagen chains (α-1, α-2, and α-3). Other identified PCM proteins included triosephosphate isomerase, transforming growth factor-β induced protein, peroxiredoxin-4, ADAM (A disintegrin and metalloproteinases) 28, and latent-transforming growth factor beta-binding protein-2. These PCM components were verified with immunohisto(cyto)chemistry for localization in the PCM region of articular cartilage. The abundance of type VI collagen in the PCM emphasizes its importance to the microenvironment of chondrocytes. Several proteins were localized in the PCM of chondrocytes for the first time and that warrants further investigation for their functions in cartilage biology.
authors.affiliation: Department of Orthopaedic Surgery, Saint Louis University, School of Medicine, 3635 Vista Avenue, Desloge Towers, DT-7, St. Louis, MO 63110, USA. zzhang4@slu.edu
authors.forename: Zijun
authors.initials: Z
authors.lastname: Zhang
authors.forename: Wu
authors.initials: W
authors.lastname: Jin
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Beckett
authors.forename: Thomas
authors.initials: T
authors.lastname: Otto
authors.forename: Berton
authors.initials: B
authors.lastname: Moed
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D002358:Cartilage, Articular / Q000378:metabolism; D019902:Chondrocytes / Q000737:chemistry / Q000378:metabolism*; D024142:Collagen Type VI / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D006801:Humans; D053779:Latent TGF-beta Binding Proteins / Q000378:metabolism; D054464:Peroxiredoxins / Q000378:metabolism*; D040901:Proteomics; D014305:Triose-Phosphate Isomerase / Q000378:metabolism*
chemical_list: D024142:Collagen Type VI; C509035:LTBP2 protein, human; D053779:Latent TGF-beta Binding Proteins; D054464:Peroxiredoxins; D051722:ADAM Proteins; C410224:ADAM28 protein, human; D014305:Triose-Phosphate Isomerase