Glutathione S-transferase omega 1-1 is a target of cytokine release inhibitory drugs and may be responsible for their effect on interleukin-1β posttranslational processing
Glutathione S-transferase omega 1-1 is a target of cytokine release inhibitory drugs and may be responsible for their effect on interleukin-1β posttranslational processing
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DOI：
10.1074/jbc.m211596200
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发表时间：
2003-05-09
影响因子：
4.8
通讯作者：
Gabel, CA
中科院分区：
文献类型：
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作者：
Laliberte, RE;Perregaux, DG;Gabel, CA
刺激诱导的人单核细胞白介素1β(IL-1β)的翻译后加工伴随着细胞内离子环境的重大变化、caspase-1的激活和细胞死亡。某些二芳基磺脲类药物可抑制这一反应，并被指定为细胞因子释放抑制药(CRID)。CRIDs阻止激活的单核细胞，使caspase-1保持非活性，并保留质膜潜伏期。[C-14]CRID的亲和标记和固定化CRID的亲和层析用于寻找潜在的作用蛋白靶点。在用含环氧化物的[C-14]CRID处理完整的人单核细胞后，谷胱甘肽S转移酶(GST)Omega 1-1被确定为首选靶点。此外，该多肽的标记与ATP诱导的IL-1β翻译后加工的不可逆抑制相关。人单核细胞提取物经CRID亲和层析后，GST Omega 1-1选择性地结合到亲和基质上，并被可溶性CRID洗脱。重组GST Omega 1-1很容易与[C-14]CrID环氧化物结合，但通过与S取代的谷胱甘肽共同孵育或通过催化中心半胱氨酸(32)突变为丙氨酸而被取消标记。高效液-质联用的多肽图谱也证明了Cys(32)是修饰的部位。尽管S-烷基谷胱甘肽不能阻止三磷酸腺苷诱导的IL-1β翻译后加工，也不能抑制[C-14]CRID掺入细胞相关的GST Omega 1-1，但谷胱甘肽-CRID加合物有效地证明了这些特性。因此，CRID抑制刺激诱导的IL-1β翻译后处理的能力可能归因于它们与GST Omega 1-1的相互作用。
Stimulus-induced posttranslational processing of human monocyte interleukin-1beta(IL-1beta) is accompanied by major changes to the intracellular ionic environment, activation of caspase-1, and cell death. Certain diarylsulfonylureas inhibit this response, and are designated cytokine release inhibitory drugs (CRIDs). CRIDs arrest activated monocytes so that caspase-1 remains inactive and plasma membrane latency is preserved. Affinity labeling with [C-14] CRIDs and affinity chromatography on immobilized CRID were used in seeking potential protein targets of their action. Following treatment of intact human monocytes with an epoxide-bearing [C-14] CRID, glutathione S-transferase (GST) Omega 1-1 was identified as a preferred target. Moreover, labeling of this polypeptide correlated with irreversible inhibition of ATP-induced IL-1beta posttranslational processing. When extracts of human monocytic cells were chromatographed on a CRID affinity column, GST Omega 1-1 bound selectively to the affinity matrix and was eluted by soluble CRID. Recombinant GST Omega 1-1 readily incorporated [C-14] CRID epoxides, but labeling was negated by co-incubation with S-substituted glutathiones or by mutagenesis of the catalytic center Cys(32) to alanine. Peptide mapping by high performance liquid chromatography-mass spectrometry also demonstrated that Cys(32) was the site of modification. Although S-alkylglutathiones did not arrest ATP-induced IL-1beta posttranslational processing or inhibit [C-14] CRID incorporation into cell-associated GST Omega 1-1, a glutathione-CRID adduct effectively demonstrated these attributes. Therefore, the ability of CRIDs to arrest stimulus-induced IL-1beta posttranslational processing may be attributable to their interaction with GST Omega 1-1.