Analysis of cartilage oligomeric matrix protein (COMP) degradation and synthesis in equine joint disease
Analysis of cartilage oligomeric matrix protein (COMP) degradation and synthesis in equine joint disease
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DOI：
10.2746/0425164054406784
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发表时间：
2005-01-01
影响因子：
2.2
通讯作者：
Sakamoto, H
中科院分区：
文献类型：
--
作者：
Arai, K;Misumi, K;Sakamoto, H
进行研究的原因：软骨寡聚基质蛋白（COMP）在软骨中含量丰富;其周转和/或降解已在各种马关节疾病中进行了研究，并已表明COMP片段化可能有助于监测此类条件。目的：确定COMP代谢是否在马骨关节炎（OA）中受损，以及COMP降解是否是代表软骨破坏的有用关节标志物。一个单克隆抗体（mAb）具有较高的亲和力降解COMP允许歧视患病关节通过量化COMP水平和fragmentation.Methods：一个单克隆抗体（克隆14 G4）产生对马软骨COMP。的NH 2-末端序列的酶切COMP片段识别14 G4进行了测定，因为是与COMP的结合效率（使用产生的COMP肽）。结果：与抗人COMP的mAb（克隆12 C4）相比，mAb 14 G4对马COMP的小片段具有更高的亲和力，其表位位于C-134和F-147之间。OA中的COMP值（平均s.d. 205.8 ± 90.9 μ g/ml）显著高于正常SF（133.1 ± 31.5 μ g/ml）。OA样品的免疫印迹，完整的COMP的比例显着降低，而较小的片段范围从75至290 kDa的较高相比，与正常SF。结论和潜在的相关性：mAb 14 G4可靠地检测COMP降解以及合成，片段分析结合定量SF可能是有用的研究马OA。
Reasons for performing study: Cartilage oligomeric matrix protein (COMP) is abundant within cartilage; its turnover and/or degradation have been investigated in various equine joint diseases and it has been suggested that COMP fragmentation might be useful for monitoring such conditions.Objectives: To determine whether COMP metabolism is compromised in equine osteoarthritis (OA) and whether COMP degradation is a useful joint marker representing cartilage destruction.Hypothesis: A monoclonal antibody (mAb) with a higher affinity for degraded COMP allows discrimination of diseased joints by quantifying COMP levels and fragmentation.Methods: A mAb (clone14G4) was generated against equine cartilage COMP. The NH2-terminal sequence of enzyme-cut COMP fragments recognised by 14G4 was determined, as was the efficiency of binding to COMP (using a generated COMP peptide). COMP concentration and fragmentation were analysed in synovial fluid (SF) from normal horses and those with OA.Results: The mAb 14G4 had a higher affinity for the smaller fragments of equine COMP, compared with a mAb (clone 12C4) generated against human COMP. The 14G4 epitope was identified as between C-134 and F-147. The COMP values in OA (mean s.d. 205.8 +/- 90.9 mug/ml) were significantly higher than in the normal (133.1 +/- 31.5 mug/ml) SF. On the immunoblots of OA sample, the proportions of intact COMP were significantly lower, while smaller fragments ranging from 75 to 290 kDa were higher compared with the normal SF.Conclusions and potential relevance: The mAb 14G4 reliably detects COMP degradation as well as synthesis, and fragmentation analysis combined with quantification in SF could be useful to study equine OA.