title: Differential protein profiling of synovial fluid from rheumatoid arthritis and osteoarthritis patients using LC-MALDI TOF/TOF.
abstract: The purpose of this study was to identify those proteins relatively more abundant in the synovial fluid (SF) of patients suffering from rheumatoid arthritis (RA) and osteoarthritis (OA) using high performance liquid chromatography coupled to mass spectrometry. 20 individual SF samples from each disease were pooled into two groups (RA and OA) to reduce the contribution of extreme individual values. Prior to the proteomic analysis, samples were immunodepleted from the top 20 most abundant plasma proteins, to enrich the lower-abundance protein fractions. Then, they were subjected to protein size fractioning and in-gel digestion, followed by reversed-phase peptide separation in a nano-LC system and subsequent peptide identification by MALDI-TOF/TOF. This strategy led to the identification of 136 different proteins in SF, which is the largest number of SF proteins described up to date by proteomics. A relative quantification of the proteins between RA and OA was carried out by spectral counting analysis. In RA, our results show a greater relative abundance of proteins related to complement activation, inflammation and the immune response, such as the major matrix metalloproteinases and several neutrophil-related proteins. In OA, we detected an increase in proteins involved in the formation and remodeling of the extracellular matrix (ECM), such as fibronectin, kininogen-1, cartilage acidic protein 1 and cartilage oligomeric matrix protein. The results obtained for MMP-1, BGH3, fibronectin and gelsolin were verified by immunoblotting analyses. Some of the novel proteins identified in this work might be relevant not only for increasing knowledge on the etiopathogenesis of RA and OA processes, but also as putative disease biomarkers, as their presence in SF is a prior step to their dilution in serum. This article is part of a Special Issue entitled: Proteomics: The clinical link.
authors.affiliation: Rheumatology Division, ProteoRed/ISCIII Proteomics Group, INIBIC-Hospital, Universitario de A Coruña, 15006-A Coruña, Spain.
authors.forename: Jesús
authors.initials: J
authors.lastname: Mateos
authors.forename: Lucía
authors.initials: L
authors.lastname: Lourido
authors.forename: Patricia
authors.initials: P
authors.lastname: Fernández-Puente
authors.forename: Valentina
authors.initials: V
authors.lastname: Calamia
authors.forename: Carlos
authors.initials: C
authors.lastname: Fernández-López
authors.forename: Natividad
authors.initials: N
authors.lastname: Oreiro
authors.forename: Cristina
authors.initials: C
authors.lastname: Ruiz-Romero
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Blanco
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001172:Arthritis, Rheumatoid / Q000378:metabolism* / Q000473:pathology; D002853:Chromatography, Liquid; D006801:Humans; D055442:Metabolome; D008875:Middle Aged; D008954:Models, Biological; D010003:Osteoarthritis / Q000378:metabolism* / Q000473:pathology; D040081:Protein Array Analysis; D020543:Proteome / Q000032:analysis / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013582:Synovial Fluid / Q000737:chemistry / Q000378:metabolism*
chemical_list: D020543:Proteome