Description of a serpin toxin in Loxosceles (Brown spider) venoms: Cloning, expression in baculovirus-infected insect cells and functional characterization

Schemczssen-Graeff, Zelinda; Justa, Hanna Câmara da; Nowatzki, Jenifer; Baldissera, Antonielle B.; Polli, Nayanne Louise Costacurta; De-Bona, Elidiana; Rossi, Izadora Volpato; Ramirez, Marcel I.; Minozzo, João Carlos; Matsubara, Fernando Hitomi; Senff‐Ribeiro, Andrea; Gremski, Luiza Helena; Veiga, Sílvio Sanches

doi:10.1016/j.ijbiomac.2021.05.129

Cited by 4 publications

(6 citation statements)

References 56 publications

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“…Finally, LSPILT induced the inhibition of melanoma cell (B16-F10) proliferation and migration, events with great participation of serine proteases. These results all together has proven that LSPILT is a molecule with a broad spectrum of inhibitory activity on serine proteases (Schemczssen- Graeff et al, 2021).…”

Section: Brown Spider Venom Moleculesmentioning

confidence: 87%

“…LSPILT is homologous to other serpins found in spiders of other genus, as well as in other arthropods. In addition, the immunoassays suggested the conservation of this class of molecules in Loxosceles gaucho, L. laeta and L. intermedia venoms (Schemczssen- Graeff et al, 2021). The expression and purification of LSPILT yielded 8.0 mg/L of culture supernatant in its soluble form and without requiring refolding.…”

Section: Brown Spider Venom Moleculesmentioning

confidence: 99%

“…However, the presence of serine protease inhibitors in the venoms of different species of Loxosceles spiders is not sufficient to prove that these molecules are in fact toxins and have harmful activities with clinical correlation to the envenoming. In this context, the inhibitory activity of plasma coagulation induced by thrombin was associated with a recombinant serpin of L. intermedia (Schemczssen- Graeff et al, 2021), which could suggest that these venom molecules may contribute with the delay of plasmatic coagulation caused by Loxosceles venom (Zanetti et al, 2002) and, consequently, increasing the hemorrhagic disturbances during loxoscelism, such as local hemorrhage observed in the cutaneous lesion (Ospedal et al, 2002). As venom components, serine protease inhibitors may also act, for instance, as protectors, preventing the proteolysis of selfproteins/toxins and then increasing the half-life of them; this contributes to the integrity of the venom, keeping its potentiality in order to be used during hunting preys, defense against predators or in accidents with humans (Dos Santos et al, 2009;Gremski et al, 2010).…”

Section: Brown Spider Venom Moleculesmentioning

confidence: 99%

“…The great advance regarding studies of serine protease inhibitors found in Loxosceles venoms emerged with the cloning and recombinant expression of serine protease inhibitor from L. intermedia in Spodoptera frugiperda cells using baculovirus technology. This molecule was called LSPILT, from Loxosceles Serine Protease Inhibitor-Like Toxin (Schemczssen- Graeff et al, 2021). Authors have shown that this protein has a molecular mass of 46 kDa, and the amino acid sequence shows the presence of the signature for the Serpin super family members.…”

Section: Brown Spider Venom Moleculesmentioning

confidence: 99%

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Prospective Use of Brown Spider Venom Toxins as Therapeutic and Biotechnological Inputs

Gremski

Matsubara

Polli

et al. 2021

Front. Mol. Biosci.

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Brown spider (genus Loxosceles) venoms are mainly composed of protein toxins used for predation and defense. Bites of these spiders most commonly produce a local dermonecrotic lesion with gravitational spread, edema and hemorrhage, which together are defined as cutaneous loxoscelism. Systemic loxoscelism, such as hematological abnormalities and renal injury, are less frequent but more lethal. Some Loxosceles venom toxins have already been isolated and extensively studied, such as phospholipases D (PLDs), which have been recombinantly expressed and were proven to reproduce toxic activities associated to the whole venom. PLDs have a notable potential to be engineered and converted in non-toxic antigens to produce a new generation of antivenoms or vaccines. PLDs also can serve as tools to discover inhibitors to be used as therapeutic agents. Other Loxosceles toxins have been identified and functionally characterized, such as hyaluronidases, allergen factor, serpin, TCTP and knottins (ICK peptides). All these toxins were produced as recombinant molecules and are biologically active molecules that can be used as tools for the potential development of chemical candidates to tackle many medical and biological threats, acting, for instance, as antitumoral, insecticides, analgesic, antigens for allergy tests and biochemical reagents for cell studies. In addition, these recombinant toxins may be useful to develop a rational therapy for loxoscelism. This review summarizes the main candidates for the development of drugs and biotechnological inputs that have been described in Brown spider venoms.

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Section: Brown Spider Venom Moleculesmentioning

confidence: 87%

Section: Brown Spider Venom Moleculesmentioning

confidence: 99%

Section: Brown Spider Venom Moleculesmentioning

confidence: 99%

Section: Brown Spider Venom Moleculesmentioning

confidence: 99%

See 2 more Smart Citations

Prospective Use of Brown Spider Venom Toxins as Therapeutic and Biotechnological Inputs

Gremski

Matsubara

Polli

et al. 2021

Front. Mol. Biosci.

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“…Insect cell expression systems are presented in various formats that have been extensively used for toxin expression, from whole insect systems, such as silkworm expression systems ( Kato et al, 2010 ), to the most commonly used, cultured insect cell lines, like the baculovirus expression vector system (BEVS) ( Deng et al, 2019 ; Justa et al, 2020 ; Schemczssen-Graeff et al, 2021 ). Baculoviruses are a group of viruses that infect insects and are harmless to humans.…”

Section: Systems For the Heterologous Expression Of Toxinsmentioning

confidence: 99%

Strategies for Heterologous Expression, Synthesis, and Purification of Animal Venom Toxins

Rivera-de-Torre

Rimbault

Jenkins

et al. 2022

Front. Bioeng. Biotechnol.

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Animal venoms are complex mixtures containing peptides and proteins known as toxins, which are responsible for the deleterious effect of envenomations. Across the animal Kingdom, toxin diversity is enormous, and the ability to understand the biochemical mechanisms governing toxicity is not only relevant for the development of better envenomation therapies, but also for exploiting toxin bioactivities for therapeutic or biotechnological purposes. Most of toxinology research has relied on obtaining the toxins from crude venoms; however, some toxins are difficult to obtain because the venomous animal is endangered, does not thrive in captivity, produces only a small amount of venom, is difficult to milk, or only produces low amounts of the toxin of interest. Heterologous expression of toxins enables the production of sufficient amounts to unlock the biotechnological potential of these bioactive proteins. Moreover, heterologous expression ensures homogeneity, avoids cross-contamination with other venom components, and circumvents the use of crude venom. Heterologous expression is also not only restricted to natural toxins, but allows for the design of toxins with special properties or can take advantage of the increasing amount of transcriptomics and genomics data, enabling the expression of dormant toxin genes. The main challenge when producing toxins is obtaining properly folded proteins with a correct disulfide pattern that ensures the activity of the toxin of interest. This review presents the strategies that can be used to express toxins in bacteria, yeast, insect cells, or mammalian cells, as well as synthetic approaches that do not involve cells, such as cell-free biosynthesis and peptide synthesis. This is accompanied by an overview of the main advantages and drawbacks of these different systems for producing toxins, as well as a discussion of the biosafety considerations that need to be made when working with highly bioactive proteins.

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Analysis of NGS data from Peruvian Loxosceles laeta spider venom gland reveals toxin diversity

Medina-Santos

Costa

Assis

et al. 2022

Comparative Biochemistry and Physiology Part D: Genomics and Pr

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Description of a serpin toxin in Loxosceles (Brown spider) venoms: Cloning, expression in baculovirus-infected insect cells and functional characterization

Cited by 4 publications

References 56 publications

Prospective Use of Brown Spider Venom Toxins as Therapeutic and Biotechnological Inputs

Prospective Use of Brown Spider Venom Toxins as Therapeutic and Biotechnological Inputs

Strategies for Heterologous Expression, Synthesis, and Purification of Animal Venom Toxins

Analysis of NGS data from Peruvian Loxosceles laeta spider venom gland reveals toxin diversity

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