Identification of prolargin expression in articular cartilage and its significance in rheumatoid arthritis pathology
| Autor: | Fabia Edathadathil, Shantikumar V. Nair, Krishnakumar N. Menon, Vinod Soman Pillai, Padmanabha Shenoy, Rameshwari R. Kundargi, Tessy Xavier, Roshini Anney Mathew, Jai Thilak |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cartilage Articular Male Osteoarthritis Cetylpyridinium chloride 01 natural sciences Biochemistry Arthritis Rheumatoid 03 medical and health sciences chemistry.chemical_compound Structural Biology Protein purification medicine Protein precipitation Humans Molecular Biology Glycoproteins chemistry.chemical_classification Extracellular Matrix Proteins Chemistry Biglycan 010401 analytical chemistry General Medicine medicine.disease Molecular biology 0104 chemical sciences Up-Regulation 030104 developmental biology Rheumatoid arthritis Urea Female Glycoprotein |
| Zdroj: | International journal of biological macromolecules. 110 |
| ISSN: | 1879-0003 |
| Popis: | Qualitative 2D gel-electrophoresis (2DE) protein profiling for osteoarthritis (OA) and rheumatoid arthritis (RA) is challenging because of selective protein loss due to discrepancies in protein precipitation methodologies. Thus, we aimed at developing qualitative protein representation from OA/RA articular cartilage without protein precipitation towards identification of clinically relevant proteins. Chondroitinase digested human articular cartilages from RA patients were subjected to protein extraction using guanidinium hydrochloride (GuHCl) or 8 M urea with 10 or 2% ASB-14-4 or 0.45 M urea with 2% ASB-14-4 with cetylpyridinium chloride (CPC). The GuHCl extract is further protein precipitated with acetone or ammonium acetate-methanol or centricon-fractionated using 100 kDa cut filters and protein precipitated using ethanol. Processed extracts were subjected to 2DE to identify protein profiles. Poor proteins representations were observed in 2D gels with protein precipitated samples compared to qualitative protein representations seen in 2D gels of 0.45 M urea and 2%ASB-14-4 extraction procedure reproducibly. The strategy circumventing protein precipitation generated qualitative 2D gels. RA vs OA gel comparison showed elevated prolargin levels in RA with biglycan levels remaining unaltered. Up regulation of prolargin in RA suggests the likelihood of an adaptive mechanism to control the increased osteoclastogenesis in RA and may have therapeutic value in controlling the disease. |
| Databáze: | OpenAIRE |
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