Establishment of a reliable method for direct proteome characterization of human articular cartilage. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Molecular and Cellular Proteomics Année : 2006

Establishment of a reliable method for direct proteome characterization of human articular cartilage.

Résumé

Articular cartilage consists mainly of extracellular matrix, mostly made of collagens and proteoglycans. These macromolecules have so far impaired its detailed two-dimensional electrophoresis-based proteomic analysis. Here, we describe a method for selective protein extraction from cartilage, which excludes proteoglycans and collagen species, thus allowing direct profiling of the protein content of cartilage by two-dimensional electrophoresis. Consistent electrophoretic patterns of more than 600 protein states were reproducibly obtained after silver staining, from 500 milligrams of human articular cartilage from joints with diverse pathologies. The extraction yield increased when the method was applied to a chondrosarcoma sample, consistent with selective extraction of cellular components. Nearly 200 of the most intensely stained protein spots were analyzed by MALDI TOF mass spectrometry after trypsin digestion. They represented 127 different proteins with diverse functions. Our method provides a rapid, efficient and pertinent alternative to previously proposed approaches for proteomic characterization of cartilage phenotypes. It will be useful at detecting protein expression patterns that relate pathophysiological processes of cartilaginous tissues such as osteoarthritis and chondrosarcoma.

Domaines

Biophysique

Dates et versions

hal-00103385 , version 1 (04-10-2006)

Identifiants

Citer

Jean-Baptiste Vincourt, Frédéric Lionneton, Gueorgui Kratassiouk, François Guillemin, Patrick Netter, et al.. Establishment of a reliable method for direct proteome characterization of human articular cartilage.. Molecular and Cellular Proteomics, 2006, 5 (10), pp.1984-1995. ⟨10.1074/mcp.T600007-MCP200⟩. ⟨hal-00103385⟩
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