Low prevalence of autoantibodies to CENP-H, -I, -K, -L, -M, -N, -T and -U in a Japanese cohort of anti-centromere positive samples
Low prevalence of autoantibodies to CENP-H, -I, -K, -L, -M, -N, -T and -U in a Japanese cohort of anti-centromere positive samples
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DOI：
10.3109/08923973.2012.733707
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发表时间：
2013-02
影响因子：
3.3
通讯作者：
A. Saito;Y. Muro;K. Sugiura;M. Akiyama
中科院分区：
文献类型：
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作者：
A. Saito;Y. Muro;K. Sugiura;M. Akiyama
目的：抗着丝粒抗体（ACA）主要靶向着丝粒区域的成分-着丝粒蛋白（CENP）-A、-B和-C.许多其他蛋白质也在间期核中围绕CENP-A核小体组装，形成间期着丝粒复合物（ICEN）。CENP-H、-I、-K、-L、-M、-N、-T和-U已被报道为ICEN的组成成分。在本研究中，我们检测了ACA与8种CENP的反应性，以研究其自身抗原性。方法：用重组CENP-B C端（Ct-CENP-B）蛋白，用免疫印迹法（WB）和酶联免疫吸附试验（ELISA）检测95例ACA患者血清。接着，通过WB用每种重组蛋白检查血清中针对8种CENP的自身抗体。此外，用计算机工具分析了各种CENP的卷曲螺旋基序和颗粒酶B（GB）切割。结果：在95份ACA阳性血清中，WB和ELISA分别有85份和93份抗Ct-CENP-B抗体阳性。在使用8种CENP的WB中，除1种血清与CENP-T弱反应外，没有血清与任何其他7种CENP反应。我们无法找到CENP的自身抗原性和卷曲螺旋形成概率或GB的潜在底物之间的任何明显关系。结论：ACA很少靶向8个CENP，而CENP-B则相反。蛋白质结构可能对CENP的自身抗原性没有主要贡献。
Objective: The constituents of the centromere region, centromere protein (CENP)-A, -B, and -C, are mainly targeted by anticentromere antibodies (ACA). Many other proteins also assemble around CENP-A nucleosomes in interphase nuclei to form the interphase centromere complex (ICEN). CENP-H, -I, -K, -L, -M, -N, -T, and -U have been reported as the constitutive components of ICEN. In this study, we examined the reactivities of ACA to the 8 CENPs for the purpose of investigating their autoantigenicity. Methods: Sera from 95 patients with ACA were tested by western blotting (WB) and enzyme-linked immunosorbent assay (ELISA) with the recombinant C-terminal of CENP-B (Ct-CENP-B). Next, the sera were examined for autoantibodies against the 8 CENPs by WB with each recombinant protein. Furthermore, the coiled-coil motifs and granzyme B (GB) cleavage for various CENPs were analyzed with computer tools. Results: Out of 95 ACA-positive sera, 85 and 93 sera were positive for anti-Ct-CENP-B antibodies in WB and in ELISA, respectively. In WB using the 8 CENPs, no sera reacted to any other 7 CENPs, except 1 serum, which reacted weakly to CENP-T. We were unable to find any obvious relationships between the autoantigenicity of CENPs and coiled-coil-forming probabilities or potential substrates for GB. Conclusion: This study demonstrates that ACA rarely target the 8 CENPs, in contrast to CENP-B. Protein structures might not contribute in a major way to the autoantigenicity of CENPs.