Diversity of astacin-like metalloproteases identified by transcriptomic analysis in Peruvian Loxosceles laeta spider venom and in vitro activity characterization

Medina-Santos, Raíssa; Guerra-Duarte, Clara; Lima, Sergio Monteiro; Costal-Oliveira, Fernanda; Aquino, Priscilla Alves de; Carmo, Anderson Oliveira do; Bonilla-Ferreyra, Cesar; Gonzalez-Kozlova, Edgar; Kalapothakis, Evanguedes; Chávez-Olórtegui, Carlos

doi:10.1016/j.biochi.2019.08.017

Cited by 16 publications

(9 citation statements)

References 44 publications

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“…The presence of metalloproteases as components of spider venom was previously detected in the venom of different Loxosceles spider species, L. intermedia , L. gaucho , L. deserta , L. laeta and L. rufescens (Feitosa et al , 1998; Young and Pincus, 2001; Da Silveira et al, 2002; Zanetti, 2002; Barbaro et. al., 2005 and Da Silveira el al., 2007) Moreover, nine possible isoforms of astacin-like metalloproteases were identified from Peruvian, L. laeta venom and validated by in silico and in vitro experiments (Medina-Santos et al, 2019). Presence of metalloproteases in the spider venom provides evidence for its significant biological activity and its conserved feature in the venom of spider species (Feitosa et al, 1998; Young and Pincus, 2001; Da Silveira et al, 2002; Zanetti, 2002 and Barbaro et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

Isolation of the Astacin-like metalloprotease coding gene (astl) and assessment of its insecticidal activity againstSpodoptera littoralisandSitophilus oryzae

Diab

Hussein

Ahmed

et al. 2019

Preprint

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14Astacin-like metalloprotease (astl) is a multi-domain metallopeptidase that has 15 protease activity against a number of organisms; including fish, frogs, birds and 16 insects. In this present investigation, the full length of astl cDNA was cloned 17 from spider species, Hasarius adansoni. Sequencing of the cloned astl cDNA 18 proved that its full length including 802 bp with 714bp open reading frame 19 encoding for 238 amino acids. The catalytic domain comprised of 489 nts was20 cloned and expressed by the yeast expression system Pichia pastoris and its 21 insecticidal activity was determined against two species of agricultural insects 22 Spodoptera littoralis (Lepidoptera:Noctuidae) and Sitophilus oryzae 23 (Coleoptera:Curculionidae). Bioassay was performed using three 2 24 concentrations (100,500 and 1000 ppm) for four days for S. littoralis and 14 25 days for S. oryzae. In addition, the astl was fused to the GNA snowdrop lectin 26 in the same frame and expressed in P. pastoris. The synergistic effect of astl 27 and GNA was examined on the S. littoralis larvae and S. oryzae adults. The 28 mortality percentages of the fused protein (Ha-astl/GNA) "1000 ppm " after 4 29 days, were 78.6%± 4.16 and 71.66% ±3.51 for first and second spodpotera 30 larval instars, respectively. While, lower mortality of the fused protein of the 31 same concentration was observed on S. oryzae adults, 49.3±2.08 %. 32 4 69pointed out that the neurotoxin protein from the Australian funnel web spider 70 Hadronyche versuta, Ѡ-hexatoxin-Hv1a was lethal to many insect species 71 when injected, while it is non-toxic through feeding. However, the Ѡ-72 hexatoxin-Hv1a/GNA fusion protein showed oral insecticidal activity against 73 insects from different orders. 74The agricultural and horticultural pest insects cause 40% loss of the crop yield 75 worldwide (Oerke et al., 1994) that is estimated by 17.7 billions dollars 76 annually (Oliveira et al., 2014). Spodoptera littoralisis one of the most 77 destructive agricultural lepidopteron insect pests that can damage many 78 economical crops such as cotton, maize, tomato and vegetables (Salama et al., 79 1970). The larvae of S. littoralis feed on the leaves, fruits, flowers, buds and 80 bolls of cotton making them useless (Bishari, 1934). Sitophilus oryzae is one of 81 the important insect pests of stored grains such as rice, wheat and their products 82 (Baloch, 1992). S. oryzae causes loss of weight, reduction in the nutritional 83 value and increasing the infection with other mites and fungi, therefore the 84 commercial value of the stored grains decreased (Madrid et al., 1990). These 85 agriculture pests are controlled by different methods such as chemical 86 insecticides which are extensively used in Egypt (Issa et al., 1984a; Issa et al., 87 1984b; and Abo-El-Ghar et al. , 1986). Therefore, resistant strains of insect 88 pests appeared in the field (Sawicki, 1986). An alternative control method that 89 used in control strategy is biological agents such as natural enemies, nuclear 90 polyhedrosi...

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Section: Discussionmentioning

confidence: 99%

Isolation of the Astacin-like metalloprotease coding gene (astl) and assessment of its insecticidal activity againstSpodoptera littoralisandSitophilus oryzae

Diab

Hussein

Ahmed

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…Metalloproteases are also expressed in high amounts in the venom gland of L. gaucho , as found by a transcriptome analysis [ 41 ]. A recent study analyzed the Peruvian L. laeta transcripts focusing on LALPs, and found 9 putative sequences coding for astacine-like metalloproteases highly similar to LALP1 from L. intermedia [ 42 ]. In addition, the authors compared the activities of Brazilian and Peruvian L. laeta venom upon fibrinogen and gelatin/collagen, and concluded that the Peruvian venom have a higher activity upon these molecules than the Brazilian one [ 42 ].…”

Section: Methodsmentioning

confidence: 99%

“…A recent study analyzed the Peruvian L. laeta transcripts focusing on LALPs, and found 9 putative sequences coding for astacine-like metalloproteases highly similar to LALP1 from L. intermedia [ 42 ]. In addition, the authors compared the activities of Brazilian and Peruvian L. laeta venom upon fibrinogen and gelatin/collagen, and concluded that the Peruvian venom have a higher activity upon these molecules than the Brazilian one [ 42 ]. A proteomic analysis of the L. intermedia crude venom using mass spectrometry also described these enzymes as components of the venom [ 43 ].…”

Section: Methodsmentioning

confidence: 99%

Brown spider venom toxins: what are the functions of astacins, serine proteases, hyaluronidases, allergens, TCTP, serpins and knottins?

Gremski

Matsubara

Justa

et al. 2021

J. Venom. Anim. Toxins incl. Trop. Dis

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Accidents caused by the bites of brown spiders (Loxosceles) generate a clinical condition that often includes a threatening necrotic skin lesion near the bite site along with a remarkable inflammatory response. Systemic disorders such as hemolysis, thrombocytopenia, and acute renal failure may occur, but are much less frequent than the local damage. It is already known that phospholipases D, highly expressed toxins in Loxosceles venom, can induce most of these injuries. However, this spider venom has a great range of toxins that probably act synergistically to enhance toxicity. The other protein classes remain poorly explored due to the difficulty in obtaining sufficient amounts of them for a thorough investigation. They include astacins (metalloproteases), serine proteases, knottins, translationally controlled tumor proteins (TCTP), hyaluronidases, allergens and serpins. It has already been shown that some of them, according to their characteristics, may participate to some extent in the development of loxoscelism. In addition, all of these toxins present potential application in several areas. The present review article summarizes information regarding some functional aspects of the protein classes listed above, discusses the directions that could be taken to materialize a comprehensive investigation on each of these toxins as well as highlights the importance of exploring the full venom repertoire.

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“…That study also showed intra-species variation in two L. laeta localities (Brazil and Peru): the Peruvian L. laeta had an additional LALP at approximately 24 kDa and more glycosylation of LALPs. The hypothesis that the Peruvian L. laeta LALPs are more enzymatic compared to the Brazilian L. laeta LALPs was confirmed by their fibrinogenolytic activity [10]. These hematological effects are typically noticeable long after the bite incident.…”

Section: Introductionmentioning

confidence: 94%

“…These hematological disturbances are most likely induced by metalloproteases, a few of which have been characterized such as Loxolysin A (20)(21)(22)(23)(24)(25)(26)(27)(28) and Loxolysin B (32)(33)(34)(35) found in L. intermedia venom [9]. Recent L. laeta venom gland transcriptomics identified multiple Loxosceles astacin-like metalloproteases (LALPs) within the 20 to 25 kDa range [10]. That study also showed intra-species variation in two L. laeta localities (Brazil and Peru): the Peruvian L. laeta had an additional LALP at approximately 24 kDa and more glycosylation of LALPs.…”

Section: Introductionmentioning

confidence: 99%

A Web of Coagulotoxicity: Failure of Antivenom to Neutralize the Destructive (Non-Clotting) Fibrinogenolytic Activity of Loxosceles and Sicarius Spider Venoms

Grashof

Zdenek

Dobson

et al. 2020

Toxins

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Envenomations are complex medical emergencies that can have a range of symptoms and sequelae. The only specific, scientifically-validated treatment for envenomation is antivenom administration, which is designed to alleviate venom effects. A paucity of efficacy testing exists for numerous antivenoms worldwide, and understanding venom effects and venom potency can help identify antivenom improvement options. Some spider venoms can produce debilitating injuries or even death, yet have been largely neglected in venom and antivenom studies because of the low venom yields. Coagulation disturbances have been particularly under studied due to difficulties in working with blood and the coagulation cascade. These circumstances have resulted in suboptimal spider bite treatment for medically significant spider genera such as Loxosceles and Sicarius. This study identifies and quantifies the anticoagulant effects produced by venoms of three Loxoscles species (L. reclusa, L. boneti, and L. laeta) and that of Sicarius terrosus. We showed that the venoms of all studied species are able to cleave the fibrinogen Aα-chain with varying degrees of potency, with L. reclusa and S. terrosus venom cleaving the Aα-chain most rapidly. Thromboelastography analysis revealed that only L. reclusa venom is able to reduce clot strength, thereby presumably causing anticoagulant effects in the patient. Using the same thromboelastography assays, antivenom efficacy tests revealed that the commonly used Loxoscles-specific SMase D recombinant based antivenom failed to neutralize the anticoagulant effects produced by Loxosceles venom. This study demonstrates the fibrinogenolytic activity of Loxosceles and Sicarius venom and the neutralization failure of Loxosceles antivenom, thus providing impetus for antivenom improvement.

show abstract

Diversity of astacin-like metalloproteases identified by transcriptomic analysis in Peruvian Loxosceles laeta spider venom and in vitro activity characterization

Cited by 16 publications

References 44 publications

Isolation of the Astacin-like metalloprotease coding gene (astl) and assessment of its insecticidal activity againstSpodoptera littoralisandSitophilus oryzae

Isolation of the Astacin-like metalloprotease coding gene (astl) and assessment of its insecticidal activity againstSpodoptera littoralisandSitophilus oryzae

Brown spider venom toxins: what are the functions of astacins, serine proteases, hyaluronidases, allergens, TCTP, serpins and knottins?

A Web of Coagulotoxicity: Failure of Antivenom to Neutralize the Destructive (Non-Clotting) Fibrinogenolytic Activity of Loxosceles and Sicarius Spider Venoms

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