Functional characterization of a fatty acyl-CoA-binding protein (ACBP) from the apicomplexan Cryptosporidium parvum.
Functional characterization of a fatty acyl-CoA-binding protein (ACBP) from the apicomplexan Cryptosporidium parvum.
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DOI：
10.1099/mic.0.28944-0
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发表时间：
2006-08
期刊：
影响因子：
1.5
通讯作者：
B. Zeng;Xiaomin Cai;G. Zhu
中科院分区：
文献类型：
--
作者：
B. Zeng;Xiaomin Cai;G. Zhu
本文报道了机会原生生物微小隐孢子虫（Cryptosporidium parvum）脂肪酰辅酶A结合蛋白（ACBP）基因的克隆和功能分析。CpACBP 1基因编码268个氨基酸的蛋白质，其比人类和动物的典型ACBP（即约90个氨基酸）大三倍。序列分析表明，CpACBP 1蛋白由N末端ACBP结构域（约90个aa）和C末端锚蛋白重复序列（约170个aa）组成。将完整的CpACBP 1 ORF工程化到麦芽糖结合蛋白融合系统中，并表达为用于功能分析的重组蛋白。酰基辅酶A结合试验清楚地表明，CpACBP 1的优选结合底物是棕榈酰辅酶A。RT-PCR、Western blotting和免疫标记分析清楚地表明CpACBP 1基因主要在胞内发育阶段表达，并且在寄生虫发育过程中表达水平增加。免疫荧光显微镜显示，CpACBP 1与寄生虫空泡膜（PVM），这意味着这种蛋白质可能参与脂质重塑的PVM，或在跨膜的脂肪酸运输。
In this paper, the identification and functional analysis of a fatty acyl-CoA-binding protein (ACBP) gene from the opportunistic protist Cryptosporidium parvum are described. The CpACBP1 gene encodes a protein of 268 aa that is three times larger than typical ACBPs (i.e. approximately 90 aa) of humans and animals. Sequence analysis indicated that the CpACBP1 protein consists of an N-terminal ACBP domain (approximately 90 aa) and a C-terminal ankyrin repeat sequence (approximately 170 aa). The entire CpACBP1 ORF was engineered into a maltose-binding protein fusion system and expressed as a recombinant protein for functional analysis. Acyl-CoA-binding assays clearly revealed that the preferred binding substrate for CpACBP1 is palmitoyl-CoA. RT-PCR, Western blotting and immunolabelling analyses clearly showed that the CpACBP1 gene is mainly expressed during the intracellular developmental stages and that the level increases during parasite development. Immunofluorescence microscopy showed that CpACBP1 is associated with the parasitophorous vacuole membrane (PVM), which implies that this protein may be involved in lipid remodelling in the PVM, or in the transport of fatty acids across the membrane.