Mitochondrial dysregulation of osteoarthritic human articular chondrocytes analyzed by proteomics: a decrease in mitochondrial superoxide dismutase points to a redox imbalance.
Mitochondria are involved in many cellular processes; mitochondrial dysfunctions have been associated with apoptosis, aging, and a number of pathological conditions, including osteoarthritis (OA). Mitochondrial proteins are attractive targets for the study of metabolism of the chondrocyte, the unique cell type present in mature cartilage, and its role in tissue degradation. Using a proteomics approach based on two-dimensional DIGE and MALDI-TOF/TOF mass spectrometric identification of mitochondria- enriched protein fractions from human articular chondrocytes, we analyzed mitochondrial protein changes that are characteristic of OA chondrocytes. A total of 73 protein forms were unambiguously identified as significantly altered in OA; 23 of them have been previously described as mitochondrial. An extensive statistical and cluster analysis of the data revealed a mitochondrial protein profile characteristic for OA. This pattern includes alterations in energy production, maintenance of mitochondrial membrane integrity, and free radical detoxification. Real time PCR, Western blot, and immunohistofluorescence assays confirmed a significant decrease of the major mitochondrial antioxidant protein manganese-superoxide dismutase (SOD2) in the superficial layer of OA cartilage. As possible outputs for this antioxidant deficiency, we found an increase of intracellular reactive oxygen species generation in OA chondrocytes and also verified an OA-dependent increase in the mitochondrial tumor necrosis factor-alpha receptor-associated protein 1 (TRAP1), a chaperone with a reported reactive oxygen species antagonist role. Our results describe the differences between the mitochondrial protein profiles of normal and OA chondrocytes, demonstrating that mitochondrial dysregulation occurs in cartilage cells during OA and highlighting redox imbalance as a key factor in OA pathogenesis.
Ruiz-Romero
Cristina
C
Osteoarticular and Aging Research Laboratory, Proteomics Unit (Nodo Asociado de Proteo-Red), Rheumatology Division, Instituto de Investigación Biomédica de A Coruña-Complejo Hospitalario Universitario A Coruña, Xubias 84, Spain.
Calamia
Valentina
V
Mateos
Jesús
J
Carreira
Vanessa
V
Martínez-Gomariz
Montserrat
M
Fernández
Mercedes
M
Blanco
Francisco J
FJ
D002358:Cartilage, Articular / Q000201:enzymology* / Q000473:pathology; D019902:Chondrocytes / Q000201:enzymology* / Q000473:pathology; D015180:Electrophoresis, Gel, Two-Dimensional; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D008928:Mitochondria / Q000201:enzymology* / Q000473:pathology*; D024101:Mitochondrial Proteins / Q000378:metabolism; D015999:Multivariate Analysis; D010003:Osteoarthritis / Q000201:enzymology* / Q000473:pathology; D010084:Oxidation-Reduction; D020543:Proteome / Q000378:metabolism; D040901:Proteomics; D017382:Reactive Oxygen Species / Q000378:metabolism; D012015:Reference Standards; D013482:Superoxide Dismutase / Q000378:metabolism*; D000099156:Superoxide Dismutase 2
D018841:HSP90 Heat-Shock Proteins; D024101:Mitochondrial Proteins; D020543:Proteome; D017382:Reactive Oxygen Species; D013482:Superoxide Dismutase; D000099156:Superoxide Dismutase 2; C092306:TRAP1 protein, human
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