Identification of citrullinated peptides in the synovial fluid of patients with rheumatoid arthritis using LC-MALDI-TOF/TOF.
Identification of citrullinated peptides in the synovial fluid of patients with rheumatoid arthritis using LC-MALDI-TOF/TOF.
复制标题
使用 LC-MALDI-TOF/TOF 鉴定类风湿关节炎患者滑液中的瓜氨酸肽
DOI：
10.1007/s10067-016-3247-4
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发表时间：
2016-09
影响因子：
3.4
通讯作者：
Li XJ
中科院分区：
文献类型：
--
作者：
Wang F;Chen FF;Gao WB;Wang HY;Zhao NW;Xu M;Gao DY;Yu W;Yan XL;Zhao JN;Li XJ
本研究的目的是探讨潜在的瓜氨酸自身抗原作为类风湿关节炎(RA)患者滑液中抗瓜氨酸蛋白抗体(ACPAs)反应的靶标。收集6例类风湿性关节炎(RA)患者和6例骨关节炎(OA)患者的SFS。用兔抗瓜氨酸抗体免疫沉淀法提取SFS中的瓜氨酸蛋白。随后进行基质辅助激光解吸/电离飞行时间/飞行时间质谱仪(MALDI-TOF/TOF)质谱仪，以发现特征中性损失，最终确定瓜氨酸自身抗原。RA SFS共鉴定出182个瓜氨酸多肽和200个瓜氨酸化位点，而OA SFS共鉴定出3个瓜氨酸多肽和4个瓜氨酸化位点。RA SFS的182个瓜氨酸多肽和OA SFS的3个瓜氨酸多肽分别来源于83和3个自身抗原。与对照组相比，除蛋白-精氨酸脱亚胺酶-2(PADI2)和蛋白-精氨酸脱亚氨酶-2(PADI4)外，其余83种自身抗原均出现瓜氨酸过度，且PADI2和PADI4的瓜氨酸结合部位不同。有趣的是，在OA对照组中发现了瓜氨酸组蛋白H3.3(H3F3A)，但在RA组中未发现。在RA SFS中发现的差异瓜氨酸蛋白表明，潜在的自身抗原是ACPAS反应的靶标，可能有助于RA补体激活和关节炎症的诱导和持续。
The objective of the study is to investigate potential citrullinated autoantigens as targets of anti-citrullinated protein antibodies (ACPAs) response in synovial fluids (SFs) of patients with rheumatoid arthritis (RA). SFs from six RA patients and six osteoarthritis (OA) patients as controls were collected. The citrullinated proteins in SFs were extracted by immunoprecipitation with rabbit anti-citrulline antibodies. Matrix-assisted laser desorption/ionization time of flight mass spectrometry/time of flight mass spectrometry (MALDI-TOF/TOF) mass spectrometry was subsequently performed to discover a characteristic neutral loss to finally determine citrullinated autoantigens. A total of 182 citrullinated peptides and 200 citrullinated sites were identified in RA SFs, while 3 citrullinated peptides and 4 citrullinated sites were identified in OA SFs. The 182 citrullinated peptides from RA SFs and the 3 citrullinated peptides from OA SFs were derived from 83 and 3 autoantigens, respectively. Eighty-three autoantigens except protein-arginine deiminase type-2 (PADI2) and protein-arginine deiminase type-2 (PADI4) were over-citrullinated compared with controls, and the citrullinated sites of PADI2 and PADI4 were different in two groups. Interestingly, citrullinated histone H3.3 (H3F3A) was found in OA controls, but not in RA groups. The differential citrullinated proteins identified in RA SFs suggested potential autoantigens were targeted for ACPAs response and might contribute to the induction and perpetuation of complement activation and joint inflammation in RA.