Characterization of Phlebotomus papatasi peritrophins, and the role of PpPer1 in Leishmania major survival in its natural vector.
Characterization of Phlebotomus papatasi peritrophins, and the role of PpPer1 in Leishmania major survival in its natural vector.
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DOI：
10.1371/journal.pntd.0002132
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发表时间：
2013
影响因子：
3.8
通讯作者：
Ramalho-Ortigao M
中科院分区：
文献类型：
--
作者：
Coutinho-Abreu IV;Sharma NK;Robles-Murguia M;Ramalho-Ortigao M
周围营养基质（PM）在血液的区隔化和许多病媒病原体的屏障中起着关键作用。为了在沙蝇中建立感染，利什曼原虫必须从内营养空间逃逸，以防止血液消化的残留物排出体外。尽管对利什曼原虫的存活起作用，但对沙蝇PM的分子组成和结构组织知之甚少。我们从木瓜白血鱼中鉴定了三种营养蛋白（PpPer1、PpPer2和PpPer3）。PpPer1和PpPer2分别显示4个和1个几丁质结合域（CBDs）。另一方面PpPer3有两个CBD，一个粘蛋白样结构域和一个具有CBD特征的推定结构域，但关键氨基酸发生了变化。时间和空间表达分析表明，PpPer1在雌性中肠经血喂养后特异性表达。PpPer2和PpPer3 mrna在中肠和后肠组成性表达，PpPer3也在马尔比氏小管中表达。PpPer2是唯一在发育阶段表达的基因。有趣的是，PpPer1和PpPer3的表达受Le的调控。主要感染。获得了重组PpPer1、PpPer2和PpPer3，并显示出与原体相似的生化特征；我们还发现PpPer1和PpPer2能够结合几丁质。PpPer1的敲低导致蛋白质减少44%，尽管对受感染的沙蝇百分比产生影响，但在48小时内导致寄生虫负荷增加39%。我们的数据表明PpPer1是papatasi P. PM的一个成分，可能参与PM作为Le屏障的作用。主要感染。为了在沙蝇媒介的中肠内成功发育，利什曼原虫必须克服媒介施加的几个障碍，包括昆虫吸食血液后在中肠内分泌的消化蛋白酶，需要逃离内营养空间，以及附着在中肠上皮上以防止血液残留物排出。沙蝇的营养基质（PM）是防止利什曼原虫在沙蝇体内形成的重要屏障，如果被困在PM内，这些寄生虫将随着血粉的残留物一起传播。尽管沙蝇PM在利什曼原虫的发展中发挥了重要作用，但目前缺乏对其分子成分的表征和对其抗利什曼原虫作用的评价。因此，我们对三种名为PpPer1、PpPer2和PpPer3的木瓜芽胞蛋白进行了分子表征。总的来说，我们证明：(1)PpPer3显示了一个假定的CBD结构域，可能已经发生了新功能化；(2)PpPer1和PpPer3基因在Le上显示了差异的基因表达。主要感染;(3) PpPer1可能是papatasi PM作为Le屏障功能的重要组成部分。主要感染。
The peritrophic matrix (PM) plays a key role in compartmentalization of the blood meal and as barrier to pathogens in many disease vectors. To establish an infection in sand flies, Leishmania must escape from the endoperitrophic space to prevent excretion with remnants of the blood meal digestion. In spite of the role played regarding Leishmania survival, little is known about sand fly PM molecular components and structural organization. We characterized three peritrophins (PpPer1, PpPer2, and PpPer3) from Phlebotomus papatasi. PpPer1 and PpPer2 display, respectively, four and one chitin-binding domains (CBDs). PpPer3 on the other hand has two CBDs, one mucin-like domain, and a putative domain with hallmarks of a CBD, but with changes in key amino acids. Temporal and spatial expression analyses show that PpPer1 is expressed specifically in the female midgut after blood feeding. PpPer2 and PpPer3 mRNAs were constitutively expressed in midgut and hindgut, with PpPer3 also being expressed in Malpighian tubules. PpPer2 was the only gene expressed in developmental stages. Interestingly, PpPer1 and PpPer3 expression are regulated by Le. major infection. Recombinant PpPer1, PpPer2 and PpPer3 were obtained and shown to display similar biochemical profiles as the native; we also show that PpPer1 and PpPer2 are able to bind chitin. Knockdown of PpPer1 led to a 44% reduction in protein, which in spite of producing an effect on the percentage of infected sand flies, resulted in a 39% increase of parasite load at 48 h. Our data suggest that PpPer1 is a component for the P. papatasi PM and likely involved in the PM role as barrier against Le. major infection. For a successful development within the midgut of the sand fly vector, Leishmania must overcome several barriers imposed by the vector that include the digestive proteases secreted within the midgut following a blood meal by the insect, the need to escape from the endoperitrophic space, and attachment to the midgut epithelia to prevent excretion with the remnants of the blood meal. The sand fly peritrophic matrix (PM) constitutes an important barrier against the establishment of Leishmania within the sand fly and if trapped within the PM these parasites will be passed along with the remnants of the blood meal. Despite the role of sand fly PM on Leishmania development, characterization of its molecular components and assessment of their roles against Leishmania are lacking. Thereby, we performed the molecular characterization of three P. papatasi peritrophins named PpPer1, PpPer2, and PpPer3. Overall, we demonstrated that: (1) PpPer3 displays a putative CBD domain that might have undertaken neo-functionalization, (2) PpPer1 and PpPer3 genes display differential gene expression upon Le. major infection; and (3) PpPer1 seems to be an important component for the function of P. papatasi PM as a barrier against Le. major infection.
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影响因子：
3.8
作者：
Coutinho-Abreu IV;Sharma NK;Robles-Murguia M;Ramalho-Ortigao M
通讯作者：
Ramalho-Ortigao M
影响因子：
4.4
作者：
Jochim RC;Teixeira CR;Laughinghouse A;Mu J;Oliveira F;Gomes RB;Elnaiem DE;Valenzuela JG
通讯作者：
Valenzuela JG
DOI：
10.1590/s0074-02762010000100001
发表时间：
2010-02-01
期刊：
Memórias do Instituto Oswaldo Cruz
影响因子：
--
作者：
Coutinho-Abreu, Iliano V;Ramalho-Ortigao, Marcelo
通讯作者：
Ramalho-Ortigao, Marcelo
影响因子：
4.4
作者：
Dillon, Rod J.;Ivens, Al C.;Churcher, Carol;Holroyd, Nancy;Quail, Michael A.;Rogers, Matthew E.;Soares, M. Bento;Bonaldo, Maria F.;Casavant, Thomas L.;Lehane, Mike J.;Bates, Paul A.
通讯作者：
Bates, Paul A.
影响因子：
4.3
作者：
Julenius, K;Molgaard, A;Brunak, S
通讯作者：
Brunak, S