Kotsyfakis Michalis, Ehret-Sabatier Laurence, Siden-Kiamos Inga, Mendoza Jaqueline, Sinden Robert E, Louis Christos
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology - Hellas, Vassilika Vouton, PO Box 1527, 71110 Heraklion, Crete, Greece.
Mol Microbiol. 2005 Jul;57(1):171-9. doi: 10.1111/j.1365-2958.2005.04664.x.
Using a proteomic approach we identified polypeptides from Anopheles gambiae and Drosophila melanogaster protein extracts that selectively bind purified Plasmodium berghei ookinetes in vitro; these were two and three distinct polypeptides, respectively, with an apparent molecular weight of about 36 kDa. Combining two-dimensional electrophoresis and MALDI-TOF (matrix-associated laser desorption ionization time of flight) mass spectrometry we determined that the polypeptides correspond to isomorphs of the annexin B11 protein of the fruit fly. When protein extracts derived from A. gambiae and D. melanogaster tissue culture cells were further fractionated, the binding activity matching the annexin protein could be localized in the fraction derived from cell membranes in both diptera. Antibody staining showed that annexin also binds to ookinetes during the invasion of the mosquito midgut. Finally, inclusion of antiannexin antisera in a mosquito blood meal impaired parasite development, suggesting a facilitating role for annexins in the infection of the mosquito by Plasmodium.
我们采用蛋白质组学方法，从冈比亚按蚊和黑腹果蝇的蛋白质提取物中鉴定出了在体外能选择性结合纯化的伯氏疟原虫动合子的多肽；这些多肽分别为两种和三种不同的多肽，表观分子量约为36 kDa。结合二维电泳和基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱分析，我们确定这些多肽对应于果蝇膜联蛋白B11的同形体。当对源自冈比亚按蚊和黑腹果蝇组织培养细胞的蛋白质提取物进行进一步分级分离时，与膜联蛋白匹配的结合活性可定位于这两种双翅目昆虫细胞膜的分级分离物中。抗体染色显示，在疟原虫侵入蚊子中肠期间，膜联蛋白也与动合子结合。最后，在蚊子血餐中加入抗膜联蛋白抗血清会损害寄生虫的发育，这表明膜联蛋白在疟原虫感染蚊子的过程中发挥促进作用。
