Biochemical evidence for a novel low molecular weight 2-5A-dependent RNase L in chronic fatigue syndrome.

Biochemical evidence for a novel low molecular weight 2-5A-dependent RNase L in chronic fatigue syndrome.
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慢性疲劳综合征中新型低分子量 2-5A 依赖性 RNase L 的生化证据。

DOI:
10.1089/jir.1997.17.377
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发表时间:
1997
期刊:
Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research
影响因子:
--
通讯作者:
Pfleiderer,W
Pfleiderer,W
中科院分区:
--
文献类型:
--
作者:
Suhadolnik,RJ;Peterson,DL;O'Brien,K;Cheney,PR;Herst,CV;Reichenbach,NL;Kon,N;Horvath,SE;Iacono,KT;Adelson,ME;DeMeirleir,K;DeBecker,P;Charubala,R;Pfleiderer,W

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该实验室之前的研究表明,在慢性疲劳综合征(CFS)中,2 ',5 ' -寡聚腺苷酸(2- 5a)合成酶/RNase L和PKR抗病毒途径的几个关键成分存在统计学意义上的失调(Suhadolnik et al. clineinfection Dis18, S96-104,1994; Suhadolnik et al.In Vivo8, 599-604,1994)。目前已有两种方法来进一步研究CFS中RNase L活性的上调。首先,用azido 2-5A光亲和探针[32P]pApAp(8-azidoA)对外周血单核细胞(PBMC)提取物进行光亲和标记,然后用抗重组人80kda RNase L的多克隆抗体进行免疫沉淀,并在变性条件下进行分析。CFS个体的一个亚群在37 kDa处仅鉴定出一个2-5A结合蛋白,而在CFS PBMC的另一个亚群和健康对照的PBMC提取物中,在80、42和37 kDa处检测到光标记/免疫反应性2-5A结合蛋白。其次,在自然条件下完成凝胶渗透高效液相色谱分析。通过poly(U)-3 ' -[32P]pCp的水解测定,健康对照PBMC提取物显示2-5A结合和2-5 a依赖的RNase L酶活性分别为80和42 kDa。CFS PBMC的一个子集含有2-5A结合蛋白,其2-5A依赖的RNase L酶活性分别为80、42和30 kDa。然而,CFS PBMC的第二个亚群含有2-5A结合和2-5A依赖的RNase L酶活性仅为30 kDa。有证据表明,CFS的RNase L酶功能障碍比以前报道的更为复杂。
Previous studies from this laboratory have demonstrated a statistically significant dysregulation in several key components of the 2′,5′-oligoadenylate (2-5A) synthetase/RNase L and PKR antiviral pathways in chronic fatigue syndrome (CFS) (Suhadolnik et al.Clin Infect Dis18, S96—104, 1994; Suhadolnik et al.In Vivo8, 599-604,1994). Two methodologies have been developed to further examine the upregulated RNase L activity in CFS. First, photoaffinity labeling of extracts of peripheral blood mononuclear cells (PBMC) with the azido 2-5A photoaffinity probe, [32P]pApAp(8-azidoA), followed by immunoprecipitation with a polyclonal antibody against recombinant, human 80-kDa RNase L and analysis under denaturing conditions. A subset of individuals with CFS was identified with only one 2-5A binding protein at 37 kDa, whereas in extracts of PBMC from a second subset of CFS PBMC and from healthy controls, photolabeled/immunoreactive 2-5A binding proteins were detected at 80, 42, and 37 kDa. Second, analytic gel permeation HPLC was completed under native conditions. Extracts of healthy control PBMC revealed 2-5A binding and 2-5 A-dependent RNase L enzyme activity at 80 and 42 kDa as determined by hydrolysis of poly(U)-3′-[32P]pCp. A subset of CFS PBMC contained 2-5A binding proteins with 2-5A-dependent RNase L enzyme activity at 80, 42, and 30 kDa. However, a second subset of CFS PBMC contained 2-5A binding and 2-5A-dependent RNase L enzyme activity only at 30 kDa. Evidence is provided indicating that the RNase L enzyme dysfunction in CFS is more complex than previously reported.
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