Self-oligomerization is essential for enhanced immunological activities of soluble recombinant calreticulin.
Self-oligomerization is essential for enhanced immunological activities of soluble recombinant calreticulin.
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自身寡聚对于可溶性重组钙网蛋白增强免疫活性至关重要
DOI：
10.1371/journal.pone.0064951
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Gao XM
中科院分区：
文献类型：
--
作者：
Huang SH;Zhao LX;Hong C;Duo CC;Guo BN;Zhang LJ;Gong Z;Xiong SD;Gong FY;Gao XM
我们最近报道了钙网蛋白（CRT），一种管腔驻留蛋白，可以在类风湿关节炎患者的血清中发现，并且重组CRT（rCRT）显示出非常强的免疫活性。我们在此进一步证明rCRT片段18-412（rCRT/18-412）、rCRT/39-272、rCRT/120-308和rCRT/120-250可以在溶液中自寡聚化，并且在体外激活巨噬细胞方面比天然CRT（nCRT，从小鼠肝脏分离）强50-100倍。我们将CRT的活性位点缩小到残基150-230，其活性也取决于二聚化。相比之下，rCRT/18-197几乎完全无活性。当通过凝胶过滤将rCRT/18-412分级分离成寡聚体和单体时，寡聚体在体外活化巨噬细胞和体内诱导特异性抗体方面保持其大部分免疫活性，而单体的活性相比之下要低得多。此外，rCRT/18-412寡聚体在与巨噬细胞结合和被巨噬细胞摄取方面比单体好得多。巨噬细胞内吞作用的抑制部分阻断了rCRT/18-412的刺激作用。我们得出结论，CRT的免疫活性位点映射在残基198-230之间，并且可溶性CRT可以在有利于其寡聚化的微环境中获得有效的免疫病理活性。
We have recently reported that calreticulin (CRT), a luminal resident protein, can be found in the sera of patients with rheumatoid arthritis and also that recombinant CRT (rCRT) exhibits extraordinarily strong immunological activities. We herein further demonstrate that rCRT fragments 18–412 (rCRT/18-412), rCRT/39-272, rCRT/120-308 and rCRT/120-250 can self-oligomerize in solution and are 50–100 fold more potent than native CRT (nCRT, isolated from mouse livers) in activating macrophages in vitro. We narrowed down the active site of CRT to residues 150–230, the activity of which also depends on dimerization. By contrast, rCRT/18-197 is almost completely inactive. When rCRT/18-412 is fractionated into oligomers and monomers by gel filtration, the oligomers maintain most of their immunological activities in terms of activating macrophages in vitro and inducing specific antibodies in vivo, while the monomers were much less active by comparison. Additionally, rCRT/18-412 oligomers are much better than monomers in binding to, and uptake by, macrophages. Inhibition of macrophage endocytosis partially blocks the stimulatory effect of rCRT/18-412. We conclude that the immunologically active site of CRT maps between residues 198–230 and that soluble CRT could acquire potent immuno-pathological activities in microenvironments favoring its oligomerization.
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影响因子：
30.5
作者：
通讯作者：
--
影响因子：
3.7
作者：
Chouquet A;Païdassi H;Ling WL;Frachet P;Houen G;Arlaud GJ;Gaboriaud C
通讯作者：
Gaboriaud C
影响因子：
5.6
作者：
Ellgaard, L;Bettendorff, P;Wüthrich, K
通讯作者：
Wüthrich, K
DOI：
10.1046/j.1432-1327.2001.02138.x
发表时间：
2001-05-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Hojrup, P;Roepstorff, P;Houen, G
通讯作者：
Houen, G
影响因子：
4.8
作者：
Kozlov, Guennadi;Pocanschi, Cosmin L.;Gehring, Kalle
通讯作者：
Gehring, Kalle