The antimalarial drug, chloroquine, interacts with lactate dehydrogenase from Plasmodium falciparum
The antimalarial drug, chloroquine, interacts with lactate dehydrogenase from Plasmodium falciparum
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DOI：
10.1016/s0166-6851(97)00095-9
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发表时间：
1997-09-01
影响因子：
1.5
通讯作者：
Foley, M
中科院分区：
文献类型：
--
作者：
Menting, JG;Tilley, L;Foley, M
我们先前已经证明，氯喹的放射性碘标记的光反应类似物，[I-125]N-（4-（4-二乙基氨基-1-甲基丁基氨基）喹啉-6-基）-4-叠氮基-2-羟基苯甲酰胺（[I-125]ASA-Q），特异性标记恶性疟原虫中表观分子量（M-r）为42和33 kDa的两种蛋白质（Foley M，Deady LW，Ng K，Cowman AF，蒂利L. J Biol Chem 1994;269：6955-6961）。我们现在报告33 kDa蛋白的鉴定。用[I-125]ASA-Q光亲和标记法从恶性疟原虫中纯化出33 kDa蛋白，以监测富集过程。纯化蛋白的N-末端序列分析显示前35个氨基酸与恶性疟原虫乳酸脱氢酶（PfLDH）完全相同。克隆并在E.大肠杆菌和重组蛋白用于生产兔抗血清。使用亲和纯化的抗PfLDH抗体的免疫沉淀证实了33 kDa CQ结合蛋白的身份。纯化的PfLDH的酶活性不受氯喹的显着影响，表明PfLDH不是CQ的直接靶标。PfLDH，但是，被证明是非常敏感的抑制自由血红素和氯喹保护这种抑制作用。(C)1997年Elsevier Science B.V.
We have previously shown that a radioiodinated photoreactive analogue of chloroquine, [I-125]N-(4-(4-diethylamino-1-methylbutylamino)quinolin-6-yl)-4-azido-2-hydroxybenzamide ([I-125]ASA-Q), specifically labels two proteins in Plasmodium falciparum with apparent molecular weights (M-r) of 42 and 33 kDa (Foley M, Deady LW, Ng K, Cowman AF, Tilley L. J Biol Chem 1994;269:6955-6961). We now report the identification of the 33 kDa protein. The 33 kDa protein was purified from Plasmodium falciparum using photoaffinity labelling with [I-125]ASA-Q to monitor the enrichment process. N-terminal sequence analysis of the purified protein revealed exact identity of the first 35 amino acids with P. falciparum lactate dehydrogenase (PfLDH). The plasmodial enzyme was cloned and expressed in E. coli and the recombinant protein used to produce a rabbit antiserum. Immunoprecipitation using affinity-purified anti-PfLDH antibodies confirmed the identity of the 33 kDa CQ-binding protein. The enzyme activity of purified PfLDH was not significantly affected by chloroquine indicating that PfLDH is not a direct target of CQ. PfLDH was, however, shown to be exquisitely sensitive to inhibition by free heme and chloroquine protected against this inhibitory effect. (C) 1997 Elsevier Science B.V.