Elucidating the Molecular Composition of Cartilage by Proteomics.
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Articular cartilage consists of chondrocytes and two major components, a collagen-rich framework and highly abundant proteoglycans. Most prior studies defining the zonal distribution of cartilage have extracted proteins with guanidine-HCl. However, an unextracted collagen-rich residual is left after extraction. In addition, the high abundance of anionic polysaccharide molecules extracted from cartilage adversely affects the chromatographic separation. In this study, we established a method for removing chondrocytes from cartilage sections with minimal extracellular matrix protein loss. The addition of surfactant to guanidine-HCl extraction buffer improved protein solubility. Ultrafiltration removed interference from polysaccharides and salts. Almost four-times more collagen peptides were extracted by the in situ trypsin digestion method. However, as expected, proteoglycans were more abundant within the guanidine-HCl extraction. These different methods were used to extract cartilage sections from different cartilage layers (superficial, intermediate, and deep), joint types (knee and hip), and disease states (healthy and osteoarthritic), and the extractions were evaluated by quantitative and qualitative proteomic analyses. The results of this study led to the identifications of the potential biomarkers of osteoarthritis (OA), OA progression, and the joint specific biomarkers.
extracellular matrix proteins
in situ trypsin digestion
multiple reaction monitoring
Extracellular Matrix Proteins
Reproducibility of Results
Published Version (Please cite this version)10.1021/acs.jproteome.5b00946
Publication InfoHsueh, Ming-Feng; Khabut, Areej; Kjellström, Sven; Önnerfjord, Patrik; & Kraus, Virginia Byers (2016). Elucidating the Molecular Composition of Cartilage by Proteomics. J Proteome Res, 15(2). pp. 374-388. 10.1021/acs.jproteome.5b00946. Retrieved from https://hdl.handle.net/10161/11580.
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Medical Instructor in the Department of Orthopaedic Surgery
Professor of Medicine
My special area of expertise is as a clinician scientist investigating osteoarthritis. Osteoarthritis is the most common form of joint disease in man and its incidence increases with age. It is a problem of increasing concern to the medical community due to the increasing longevity of the population. Trained as a molecular biologist and a Rheumatologist, I endeavor to study this disease from bedside to bench. The work in this laboratory focuses on osteoarthritis and deals w
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