Tabushi Yoko, Nakanishi Toyofumi, Takeuchi Tohru, Nakajima Mikio, Ueda Kazuhito, Kotani Takuya, Makino Shigeki, Shimizu Akira, Hanafusa Toshiaki, Takubo Takayuki
Department of Clinical Pathology, Osaka Medical College, Osaka, Japan.
Ann Clin Biochem. 2008 Jul;45(Pt 4):413-7. doi: 10.1258/acb.2007.007205.
Rheumatoid arthritis (RA) is the most common inflammatory joint disease. The aetiology of RA remains unknown, but autoimmune responses are considered to play an important role in the disease pathophysiology. Currently available data suggests that the process of diagnosing RA may benefit from testing for anticyclic citrullinated peptides. Identification of the presence of citrullinated proteins in rheumatoid synovial fluids is important for the elucidation of the aetiology of RA as well as in the differential diagnosis of rheumatic-related diseases.
A proteomics-based approach using electrophoresis/mass spectrometry was applied to identify the citrullinated proteins in synovial fluids from patients with RA. Synovial fluids from patients with RA were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis to detect the citrullinated proteins. Identification bands were then subjected to mass spectrometry.
Three proteins - citrullinated fibrinogen, citrullinated fibronectin and citrullinated vimentin - in synovial fluids from RA patients were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.
Proteomics-based analysis can be used to detect citrullinated proteins in synovial fluids from RA patients.
类风湿关节炎(RA)是最常见的炎性关节疾病。RA的病因尚不清楚,但自身免疫反应被认为在该疾病的病理生理过程中起重要作用。目前可得的数据表明,RA的诊断过程可能受益于抗环瓜氨酸肽检测。类风湿滑膜液中瓜氨酸化蛋白的鉴定对于阐明RA的病因以及风湿相关疾病的鉴别诊断都很重要。
采用基于蛋白质组学的电泳/质谱方法来鉴定RA患者滑膜液中的瓜氨酸化蛋白。对RA患者的滑膜液进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和蛋白质印迹分析,以检测瓜氨酸化蛋白。然后对鉴定条带进行质谱分析。
通过基质辅助激光解吸电离飞行时间质谱法鉴定出RA患者滑膜液中的三种蛋白——瓜氨酸化纤维蛋白原、瓜氨酸化纤连蛋白和瓜氨酸化波形蛋白。
基于蛋白质组学的分析可用于检测RA患者滑膜液中的瓜氨酸化蛋白。