Identification, cDNA cloning and heterologous expression of a hyaluronidase from the tarantula Brachypelma vagans venom

Clement, Herlinda; Olvera, Alejandro; Rodríguez, Mabel; Zamudio, Fernando Z.; Palomares, Laura A.; Possani, Lourival D.; Odell, George V.; Alagón, Alejandro; Sánchez‐López, Rosana

doi:10.1016/j.toxicon.2012.08.018

Cited by 20 publications

(27 citation statements)

References 29 publications

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“…Cysteines and disulfide bonds were highly conserved in the catalytic domain; the C 172 -C 215 disulfide bond (Fig. 2, marked by the red arrow) appeared to be exclusive to arachnidic hyaluronidases and, as Clement et al suggested, may reinforce the stability of the catalytic site [41]. Cysteine residues in the EGF-like domain (from C 331 to the C-terminus) were also conserved.…”

Section: Discussionmentioning

confidence: 88%

“…Although there are several potential N-glycosylation sites for each sequence aligned, they were not well conserved among each other (Fig. 2, light blue squares) [65], [54], [52], [41], [43]. GPP analysis showed that N-glycosylation sites were not conserved even between TsHyal-1 and TsHyal-2 isoforms.…”

Section: Discussionmentioning

confidence: 96%

“…Ts hyaluronidase sequences were aligned with known hyaluronidase sequences from diverse organisms, including Mesobuthus martensii (GenBank: ACY69673; [52]), Loxosceles intermedia (GenBank: AGH25912; [42]), Brachypelma vagans (GenBank: AFS33217; [41]), Polybia paulista (GenBank: ADL 09135; [43]), Apis mellifera (UniProtKB/Swiss-Prot: Q08169; [53]), and Homo sapiens (UniProtKB/Swiss-Prot: Q12794; [54]). A multiple alignment was performed using the algorithm Clustal W in MEGA 5.1 software [55].…”

Section: Methodsmentioning

confidence: 99%

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Molecular, Immunological, and Biological Characterization of Tityus serrulatus Venom Hyaluronidase: New Insights into Its Role in Envenomation

et al. 2014

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BackgroundScorpionism is a public health problem in Brazil, and Tityus serrulatus (Ts) is primarily responsible for severe accidents. The main toxic components of Ts venom are low-molecular-weight neurotoxins; however, the venom also contains poorly characterized high-molecular-weight enzymes. Hyaluronidase is one such enzyme that has been poorly characterized.Methods and principal findingsWe examined clones from a cDNA library of the Ts venom gland and described two novel isoforms of hyaluronidase, TsHyal-1 and TsHyal-2. The isoforms are 83% identical, and alignment of their predicted amino acid sequences with other hyaluronidases showed conserved residues between evolutionarily distant organisms. We performed gel filtration followed by reversed-phase chromatography to purify native hyaluronidase from Ts venom. Purified native Ts hyaluronidase was used to produce anti-hyaluronidase serum in rabbits. As little as 0.94 µl of anti-hyaluronidase serum neutralized 1 LD50 (13.2 µg) of Ts venom hyaluronidase activity in vitro. In vivo neutralization assays showed that 121.6 µl of anti-hyaluronidase serum inhibited mouse death 100%, whereas 60.8 µl and 15.2 µl of serum delayed mouse death. Inhibition of death was also achieved by using the hyaluronidase pharmacological inhibitor aristolochic acid. Addition of native Ts hyaluronidase (0.418 µg) to pre-neutralized Ts venom (13.2 µg venom+0.94 µl anti-hyaluronidase serum) reversed mouse survival. We used the SPOT method to map TsHyal-1 and TsHyal-2 epitopes. More peptides were recognized by anti-hyaluronidase serum in TsHyal-1 than in TsHyal-2. Epitopes common to both isoforms included active site residues.ConclusionsHyaluronidase inhibition and immunoneutralization reduced the toxic effects of Ts venom. Our results have implications in scorpionism therapy and challenge the notion that only neurotoxins are important to the envenoming process.

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Section: Discussionmentioning

confidence: 88%

Section: Discussionmentioning

confidence: 96%

Section: Methodsmentioning

confidence: 99%

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Molecular, Immunological, and Biological Characterization of Tityus serrulatus Venom Hyaluronidase: New Insights into Its Role in Envenomation

et al. 2014

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“…AnHyal showed specificity to the hyaluronic acid substrate, like the one produced by Brachypelma vagans [55] and H. partita [38]. Conversely, hyaluronidase from L. intermedia [44], V. dubius [39], and several other species of spiders [7], were also able to degrade chondroitin sulfate.…”

Section: Accepted Manuscriptmentioning

confidence: 97%

Biochemical and structural characterization of a protein complex containing a hyaluronidase and a CRISP-like protein isolated from the venom of the spider Acanthoscurria natalensis

Barth

Mandacaru

Charneau

et al. 2019

Journal of Proteomics

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“…vezes mais ativa que a hialuronidase isolada de serpentes, como a de Crotalus durissus terrificus (PESSINI et al, 2001;BORDON et al, 2012 (FERRER et al, 2013;SKOV et al, 2006;GMACHL;KREIL et al, 1993), leveduras (REITINGER et al, 2008JIN et al, 2014), plantas (JUNG et al 2010), células de inseto (CLEMENT et al, 2012;SOLDATOVA et al, 2007;HOFINGER et al, 2007;NG et al, 2005;SOLDATOVA et al, 1998) (KING et al, 1996), no peixe pedra Synanceja horrida (NG et al, 2005)…”

Section: Hialuronidases Da Peçonha De Escorpiãounclassified

Clonagem e expressão heteróloga da hialuronidase e/ou novas toxinas obtidas a partir do transcriptoma da glândula da peçonha de Tityus serrulatus

Amorim¹

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Agradeço a Deus por determinar a mim esse destino e colocar tantas pessoas especiais na minha vida. Agradeço à minha família por todo apoio e compreensão nos momentos em que estive ausente. Esta conquista é dedicada em memória ao meu querido pai Antonio, que era o meu maior incentivador para que eu seguisse esse caminho. Agradeço também às mulheres da minha vida, minha mãe Valeria e minha irmã Vanessa, por toda a força que me deram quando mais precisei e se não fosse isso eu não teria chegado até aqui. O que sou hoje é graças a vocês. É o amor de vocês que me dá forças! Amo vocês! Ao Lucas, meu amado esposo, meu orgulho. Obrigada por ser meu companheiro em todas as jornadas e ajudar no meu crescimento pessoal e profissional. Há pessoas que entram na nossa vida para nos mostrar o melhor de nós e nos fazer bem. A felicidade só é real quando compartilhada e juntos nós vamos longe. Te amo! À Profa. Dr a. Eliane Candiani Arantes Braga por me aceitar no grupo de pesquisa e por embarcar no desafio desse projeto. Ser sua aluna foi um privilégio e eu sou eternamente grata por todo aprendizado e conhecimento que você permitiu que eu adquirisse. Obrigada por compreender todos os momentos em que precisei me ausentar e por nos incentivar a pensar grande. Parte do profissional que eu sou, e serei, vem do exemplo que obtive de você. Eu não poderia ter orientadora melhor que você. Obrigada por tudo! À Profa. Dra. Simone Kashima pela coorientação e conhecimento compartilhado durante o doutorado. Muito obrigada por me receber em seu laboratório e pelo direcionamento do trabalho em uma fase tão importante. Agradeço à amiga Dra. Virginia Mara de Deus Wagatsuma por toda a ajuda nos experimentos da biblioteca de cDNA e todas as aventuras experimentais que realizamos na busca da sequência da hialuronidase. Obrigada pela sua disponibilidade em me ajudar, sempre disposta e otimista. Não encontrei a sequência, mas ganhei uma amiga!

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Identification, cDNA cloning and heterologous expression of a hyaluronidase from the tarantula Brachypelma vagans venom

Cited by 20 publications

References 29 publications

Molecular, Immunological, and Biological Characterization of Tityus serrulatus Venom Hyaluronidase: New Insights into Its Role in Envenomation

Molecular, Immunological, and Biological Characterization of Tityus serrulatus Venom Hyaluronidase: New Insights into Its Role in Envenomation

Biochemical and structural characterization of a protein complex containing a hyaluronidase and a CRISP-like protein isolated from the venom of the spider Acanthoscurria natalensis

Clonagem e expressão heteróloga da hialuronidase e/ou novas toxinas obtidas a partir do transcriptoma da glândula da peçonha de Tityus serrulatus

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