Conformational co-dependence between Plasmodium berghei LCCL proteins promotes complex formation and stability.
Conformational co-dependence between Plasmodium berghei LCCL proteins promotes complex formation and stability.
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DOI：
10.1016/j.molbiopara.2012.07.007
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发表时间：
2012-10
影响因子：
1.5
通讯作者：
Dessens JT
中科院分区：
文献类型：
--
作者：
Saeed S;Tremp AZ;Dessens JT
本文通过杂交 PbLAP1-KO 和 PbLAP3/GFP 寄生虫系，鉴定了伯氏疟原虫 LCCL 蛋白之间的构象相互依赖性。 ► 伯氏疟原虫中似乎不存在 LCCL 蛋白的共依赖性表达。 ► 然而，伯氏疟原虫 LCCL 蛋白在构象上是相互依赖的。 ► 该机制促进LCCL蛋白复合物的形成和稳定性。 ► 构象相互依赖可能是相互依赖表达背后的机制。疟原虫表达一个保守的 LCCL 凝集素粘附样结构域蛋白 (LAP) 家族，该蛋白在子孢子传播中具有重要功能。在恶性疟原虫中，所有六个家族成员均在配子细胞中表达并形成多蛋白复合物。有趣的是，敲除恶性疟原虫 LCCL 蛋白会对其他家族成员的蛋白表达产生不利影响，但不会影响 mRNA 水平，这种现象称为共依赖性表达。在这里，我们通过将 PbLAP1 无效突变寄生虫与表达 GFP 标记的 PbLAP3 的寄生虫系（在配子细胞中显示出强荧光）杂交来研究伯氏疟原虫的这一现象。选定并验证的双突变体显示 PbLAP3::GFP 的正常合成和亚细胞定位。然而，在没有 PbLAP1 存在的情况下，基于 GFP 的荧光显着降低，表明 PbLAP1 和 PbLAP3 相互作用。此外，PbLAP1 的缺失会显着缩短 PbLAP3 的半衰期，这与错误折叠的情况一致。这些发现揭示了构象相互依赖的潜在机制，促进功能性 LCCL 蛋白复合物的组装和稳定性。
This paper identifies conformational codependence between Plasmodium berghei LCCL proteins by crossing PbLAP1-KO and PbLAP3/GFP parasite lines. ► Co-dependent expression of LCCL proteins appears absent in Plasmodium berghei. ► However, P. berghei LCCL proteins are conformationally co-dependent. ► This mechanism promotes LCCL protein complex formation and stability. ► Conformational co-dependence could be the mechanism behind co-dependent expression. Malaria parasites express a conserved family of LCCL-lectin adhesive-like domain proteins (LAPs) that have essential functions in sporozoite transmission. In Plasmodium falciparum all six family members are expressed in gametocytes and form a multi-protein complex. Intriguingly, knockout of P. falciparum LCCL proteins adversely affects expression of other family members at protein, but not at mRNA level, a phenomenon termed co-dependent expression. Here, we investigate this in Plasmodium berghei by crossing a PbLAP1 null mutant parasite with a parasite line expressing GFP-tagged PbLAP3 that displays strong fluorescence in gametocytes. Selected and validated double mutants show normal synthesis and subcellular localization of PbLAP3::GFP. However, GFP-based fluorescence is dramatically reduced without PbLAP1 present, indicating that PbLAP1 and PbLAP3 interact. Moreover, absence of PbLAP1 markedly reduces the half-life of PbLAP3, consistent with a scenario of misfolding. These findings unveil a potential mechanism of conformational interdependence that facilitates assembly and stability of the functional LCCL protein complex.
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