A Rhipicephalus (Boophilus) microplus cathepsin with dual peptidase and antimicrobial activity

Oldiges, Daiane P.; Parizi, Luís Fernando; Zimmer, Karine Rigon; Lorenzini, Daniel M.; Seixas, Adriana; Masuda, Aoi; Vaz, Itabajara da Silva; Termignoni, Carlos

doi:10.1016/j.ijpara.2012.04.013

Cited by 19 publications

(13 citation statements)

References 78 publications

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“…Cysteine proteases are important constituents of the immune response of R. microplus and participate in vitellin degradation [33,34]. The unigene MPOAH54TR had high similarity to a cysteine protease, longipain.…”

Section: Resultsmentioning

confidence: 99%

The ovarian transcriptome of the cattle tick, Rhipicephalus (Boophilus) microplus, feeding upon a bovine host infected with Babesia bovis

et al. 2013

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BackgroundCattle babesiosis is a tick-borne disease of cattle with the most severe form of the disease caused by the apicomplexan, Babesia bovis. Babesiosis is transmitted to cattle through the bite of infected cattle ticks of the genus Rhipicephalus. The most prevalent species is Rhipicephalus (Boophilus) microplus, which is distributed throughout the tropical and subtropical countries of the world. The transmission of B. bovis is transovarian and a previous study of the R. microplus ovarian proteome identified several R. microplus proteins that were differentially expressed in response to infection. Through various approaches, we studied the reaction of the R. microplus ovarian transcriptome in response to infection by B. bovis.MethodsA group of ticks were allowed to feed on a B. bovis-infected splenectomized calf while a second group fed on an uninfected splenectomized control calf. RNA was purified from dissected adult female ovaries of both infected and uninfected ticks and a subtracted B. bovis-infected cDNA library was synthesized, subtracting with the uninfected ovarian RNA. Four thousand ESTs were sequenced from the ovary subtracted library and annotated.ResultsThe subtracted library dataset assembled into 727 unique contigs and 2,161 singletons for a total of 2,888 unigenes, Microarray experiments designed to detect B. bovis-induced gene expression changes indicated at least 15 transcripts were expressed at a higher level in ovaries from ticks feeding upon the B. bovis-infected calf as compared with ovaries from ticks feeding on an uninfected calf. We did not detect any transcripts from these microarray experiments that were expressed at a lower level in the infected ovaries compared with the uninfected ovaries. Using the technique called serial analysis of gene expression, 41 ovarian transcripts from infected ticks were differentially expressed when compared with transcripts of controls.ConclusionCollectively, our experimental approaches provide the first comprehensive profile of the R. microplus ovarian transcriptome responding to infection by B. bovis. This dataset should prove useful in molecular studies of host-pathogen interactions between this tick and its apicomplexan parasite.

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Section: Resultsmentioning

confidence: 99%

The ovarian transcriptome of the cattle tick, Rhipicephalus (Boophilus) microplus, feeding upon a bovine host infected with Babesia bovis

et al. 2013

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“…Although VTDCE has been classified as a cathepsin L-like cysteine [ 76 ], a very low similarity was found between its deduced amino acid sequence (AFK78425.1) and any other cysteine endopeptidase. On the other hand, phylogenetic sequence analysis revealed that VTDCE is similar to some tick antimicrobial peptides [ 78 ]. Moreover, the presence of VTDCE significantly inhibits Staphylococcus epidermidis growth after a period of 24 h. This is the first arthropod protease to be reported as an antimicrobial that is not correlated with its peptidase activity [ 78 ].…”

Section: Proteases From Haematophagous Arthropod Vectorsmentioning

confidence: 99%

Proteases of haematophagous arthropod vectors are involved in blood-feeding, yolk formation and immunity - a review

et al. 2017

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Ticks, triatomines, mosquitoes and sand flies comprise a large number of haematophagous arthropods considered vectors of human infectious diseases. While consuming blood to obtain the nutrients necessary to carry on life functions, these insects can transmit pathogenic microorganisms to the vertebrate host. Among the molecules related to the blood-feeding habit, proteases play an essential role. In this review, we provide a panorama of proteases from arthropod vectors involved in haematophagy, in digestion, in egg development and in immunity. As these molecules act in central biological processes, proteases from haematophagous vectors of infectious diseases may influence vector competence to transmit pathogens to their prey, and thus could be valuable targets for vectorial control.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-017-2005-z) contains supplementary material, which is available to authorized users.

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“…This result shows that polyclonal antibodies were able to block the inhibitory activity of rRmS-3 by approximately 66%, of rRmS-6 by 50% and of rRmS-17 by 85% (Table 3). A non-related serpin anti-serum was used as control (anti-VTDCE) (Oldiges et al, 2012) and did not affect serpin inhibitory activity(Fig. S1).…”

Section: Resultsmentioning

confidence: 99%

The putative role of Rhipicephalus microplus salivary serpins in the tick-host relationship

Tirloni

Kim

Coutinho

et al. 2016

Insect Biochemistry and Molecular Biology

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Inflammation and haemostasis are part of the host's first line of defense to tick feeding. These systems are in part serine protease mediated and are tightly controlled by their endogenous inhibitors, in the serpin superfamily (serine protease inhibitors). From this perspective ticks are thought to use serpins to evade host defenses during feeding. The cattle tick Rhipicephalus microplus encodes at least 24 serpins, of which RmS-3, RmS-6, and RmS-17 were previously identified in saliva of this tick. In this study, we screened inhibitor functions of these three saliva serpins against a panel of 16 proteases across the mammalian defense pathway. Our data confirm that Pichia pastorisexpressed rRmS-3, rRmS-6, and rRmS-17 are likely inhibitors of pro-inflammatory and pro-coagulant proteases. We show that rRmS-3 inhibited chymotrypsin and cathepsin G with stoichiometry of inhibition (SI) indices of 1.8 and 2.0, and pancreatic elastase with SI higher than 10. Likewise, rRmS-6 inhibited trypsin with SI of 2.6, chymotrypsin, factor Xa, factor XIa, and plasmin with SI higher than 10, while rRmS-17 inhibited trypsin, cathepsin G, chymotrypsin, plasmin, and factor XIa with SI of 1.6, 2.6, 2.7, 3.4, and 9.0, respectively. Additionally, we observed the formation of irreversible complexes between rRmS-3 and chymotrypsin, rRmS-6/rRmS-17 and trypsin, and rRmS-3/rRmS-17 and cathepsin G, which is consistent with typical mechanism of inhibitory serpins. In blood clotting assays, rRmS-17 delayed plasma clotting by 60 s in recalcification time assay, while rRmS-3 and rRmS-6 did not have any effect. Consistent with inhibitor function profiling data, 2.0 µM rRmS-3 and rRmS-17 inhibited cathepsin G-activated platelet aggregation in a dose-responsive manner by up to 96% and 95% respectively. Of significant interest, polyclonal antibodies blocked inhibitory functions of the three serpins. Also notable, antibodies to Amblyomma americanum, Ixodes scapularis, and R. sanguineus tick saliva proteins cross-reacted with the three R. microplus saliva serpins, suggesting the potential of these proteins as candidates for universal anti-tick vaccines.

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A Rhipicephalus (Boophilus) microplus cathepsin with dual peptidase and antimicrobial activity

Cited by 19 publications

References 78 publications

The ovarian transcriptome of the cattle tick, Rhipicephalus (Boophilus) microplus, feeding upon a bovine host infected with Babesia bovis

The ovarian transcriptome of the cattle tick, Rhipicephalus (Boophilus) microplus, feeding upon a bovine host infected with Babesia bovis

Proteases of haematophagous arthropod vectors are involved in blood-feeding, yolk formation and immunity - a review

The putative role of Rhipicephalus microplus salivary serpins in the tick-host relationship

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