Department of Parasitology, Faculty of Science, Charles University, Vinicna 7, Prague 2, 128 44, Czech Republic.
Department of Parasitology, Faculty of Science, Charles University, Vinicna 7, Prague 2, 128 44, Czech Republic.
Insect Biochem Mol Biol. 2018 Nov;102:67-74. doi: 10.1016/j.ibmb.2018.09.010. Epub 2018 Sep 28.
Current knowledge of sand fly salivary components has been based solely on Lutzomyia and Phlebotomus species which feed mainly on mammals; their hyaluronidases and apyrases were demonstrated to significantly affect blood meal intake and transmission of vector-borne pathogens. Members of the third sand fly genus Sergentomyia preferentially feed on reptiles but some of them are considered as Leishmania and arboviruses vectors; however, nothing is known about their salivary components that might be relevant for pathogens transmission. Here, marked hyaluronidase and apyrase activities were demonstrated in the saliva of a Sergentomyia schwetzi colony maintained on geckos. Hyaluronidase of S. schwetzi cleaved hyaluronan as the prominent substrate, and was active over a broad pH range from 4.0 to 8.0, with a sharp peak at pH 5.0. SDS PAGE zymography demonstrated the monomeric character of the enzyme, which remained active in reducing conditions. The apparent molecular weight of 43 kDa was substantially lower than in any sand fly species tested so far and may indicate relatively low grade of the glycosylation of the enzyme. The apyrase of S. schwetzi was typical strictly Ca2+ dependent Cimex-family apyrase. It was active over a pH range from 6.5 to 9.0, with a peak of activity at pH 8.5, and had an ATPase/ADPase ratio of 0.9. The apyrase activity increased during the first 3 days post-emergence, then reached a plateau and remained relatively constant until day 8. In comparison with a majority of Phlebotomus and Lutzomyia species tested to date, both the hyaluronidase and apyrase activities of S. schwetzi were relatively low, which may reflect an adaptation of this sand fly to blood feeding on non-mammalian hosts.
目前对沙蝇唾液成分的了解仅基于主要以哺乳动物为食的 Lutzomyia 和 Phlebotomus 物种;它们的透明质酸酶和 ADP 酶已被证明显著影响了血液摄入和媒介传播病原体。第三类沙蝇属 Sergentomyia 的成员更喜欢以爬行动物为食,但其中一些被认为是利什曼原虫和虫媒病毒的媒介;然而,对于可能与病原体传播相关的它们的唾液成分,我们一无所知。在这里,在以壁虎为食的 Sergentomyia schwetzi 种群的唾液中,明显显示出透明质酸酶和 ADP 酶的活性。S. schwetzi 的透明质酸酶以透明质酸为主要底物进行切割,在 pH 值为 4.0 至 8.0 的较宽范围内具有活性,在 pH 值 5.0 时具有明显的峰值。SDS PAGE 酶谱法证明了该酶的单体特性,该酶在还原条件下仍保持活性。43 kDa 的表观分子量明显低于迄今为止测试的任何沙蝇种,这可能表明该酶的糖基化程度相对较低。S. schwetzi 的 ADP 酶是典型的严格依赖 Ca2+的 Cimex 家族 ADP 酶。它在 pH 值为 6.5 至 9.0 的范围内具有活性,在 pH 值 8.5 时具有活性峰值,并且 ATPase/ADPase 比值为 0.9。在出蛰后的前 3 天,APY 酶活性增加,然后达到平台期,并在第 8 天之前保持相对稳定。与迄今为止测试的大多数 Phlebotomus 和 Lutzomyia 物种相比,S. schwetzi 的透明质酸酶和 ADP 酶活性相对较低,这可能反映了这种沙蝇对非哺乳动物宿主血液的适应性。