Functional and structural characterization of an ecotin-like serine protease inhibitor from Trypanosoma cruzi

Garcia, Felipe Baena; Cabral, Aline Diniz; Fuhlendorf, Max Mario; Cruz, Geomar F.; Santos, Juliete Vitorino dos; Ferreira, Graziele Cristina; Rezende, Bernard Robin Carneiro de; Santana, Carla Moreira; Puzer, Luciano; Sasaki, Sergio D.; Garcia, Wânius; Sperança, Márcia Aparecida

doi:10.1016/j.ijbiomac.2020.02.186

Cited by 6 publications

(13 citation statements)

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“…Ecotin was first described in E. coli as a periplasmic protease inhibitor that exhibits a broad specificity toward exogenous serine proteases including trypsin, chymotrypsin, factor Xa, NE, kallikrein, urokinase and factor XII, but not against metallo-, aspartyl and sulfhydryl proteases, or its own proteases [28][29][30][31][32]. Ecotin homologs have since been found in more than 300 organisms, predominantly in Gram-negative bacterial pathogens such as Pseudomonas aeruginosa, 33], but also in two eukaryotic protozoan parasites within the Trypanosomatidae genus (Leishmania major and Trypanosoma cruzi [12,34,35]), and in some plant pathogens e. g. Pantoea citrea [31]. In the latter case, ecotin was found to be less potent in binding to NE when compared to the E. coli homolog, suggesting that the protein may have evolved to recognize alternate proteases specific to its host [31].…”

Section: Introductionmentioning

confidence: 99%

Characterization of ecotin homologs from Campylobacter rectus and Campylobacter showae

et al. 2020

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Ecotin, first described in Escherichia coli, is a potent inhibitor of a broad range of serine proteases including those typically released by the innate immune system such as neutrophil elastase (NE). Here we describe the identification of ecotin orthologs in various Campylobacter species, including Campylobacter rectus and Campylobacter showae residing in the oral cavity and implicated in the development and progression of periodontal disease in humans. To investigate the function of these ecotins in vitro, the orthologs from C. rectus and C. showae were recombinantly expressed and purified from E. coli. Using CmeA degradation/protection assays, fluorescence resonance energy transfer and NE activity assays, we found that ecotins from C. rectus and C. showae inhibit NE, factor Xa and trypsin, but not the Campylobacter jejuni serine protease HtrA or its ortholog in E. coli, DegP. To further evaluate ecotin function in vivo, an E. coli ecotin-deficient mutant was complemented with the C. rectus and C. showae homologs. Using a neutrophil killing assay, we demonstrate that the low survival rate of the E. coli ecotin-deficient mutant can be rescued upon expression of ecotins from C. rectus and C. showae. In addition, the C. rectus and C. showae ecotins partially compensate for loss of N-glycosylation and increased protease susceptibility in the related pathogen, Campylobacter jejuni, thus implicating a similar role for these proteins in the native host to cope with the protease-rich environment of the oral cavity.

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Section: Introductionmentioning

confidence: 99%

Characterization of ecotin homologs from Campylobacter rectus and Campylobacter showae

et al. 2020

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“…For example, Pseudomonas fuscovaginae, Burkholderia glumae, and Burkholderia plantarii are important pathogens of rice, while Ganoderma boninense and Gibbsiella quercinecans cause stem rot of oil palm and acute oak decline, respectively (Brady et al, 2016;Isha et al, 2020). The genomes of several Eukaryotes also encode ecotin orthologs, especially within the phylum Euglenozoa, which contains the insectborne parasitic genera Trypanosoma and Leishmania (Peña et al, 2017;Verma et al, 2017Verma et al, , 2018Garcia et al, 2020;Levy et al, 2021). These genera contain species such as Trypanosoma cruzi, which causes Chagas disease (Brener, 1973), Leishmania major causing zoonotic cutaneous leishmaniasis (El-On et al, 1984), and Leishmania donovani the causative agent of visceral leishmaniasis, traditionally known as kala-azar ("black fever"; van Griensven and Diro, 2012).…”

Section: Taxonomic Distribution Of Ecotinmentioning

confidence: 99%

“…Ecotin has been studied for nearly four decades ( Chung et al, 1983 ), and more than 600 protein homologs have since been discovered across the Bacterial and Eukaryotic kingdoms ( Figure 2 ; Eschenlauer et al, 2009 ; Ireland et al, 2014 ; Nagy et al, 2019 ; Garcia et al, 2020 ). Ecotin sequences diverge along the major taxonomic lineages, indicating that homologs were present in the ancestors of Proteobacteria ( Figure 2C ).…”

Section: Taxonomic Distribution Of Ecotinmentioning

confidence: 99%

“…The genomes of several Eukaryotes also encode ecotin orthologs, especially within the phylum Euglenozoa , which contains the insect-borne parasitic genera Trypanosoma and Leishmania ( Peña et al, 2017 ; Verma et al, 2017 , 2018 ; Garcia et al, 2020 ; Levy et al, 2021 ). These genera contain species such as Trypanosoma cruzi , which causes Chagas disease ( Brener, 1973 ), Leishmania major causing zoonotic cutaneous leishmaniasis ( El-On et al, 1984 ), and Leishmania donovani the causative agent of visceral leishmaniasis, traditionally known as kala-azar (“black fever”; van Griensven and Diro, 2012 ).…”

Section: Taxonomic Distribution Of Ecotinmentioning

confidence: 99%

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Ecotin: A versatile protease inhibitor of bacteria and eukaryotes

Meyer

Carlier

2023

Front. Microbiol.

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Serine protease inhibitors are a large family of proteins involved in important pathways and processes, such as inflammatory responses and blood clotting. Most are characterized by a precise mode of action, thereby targeting a narrow range of protease substrates. However, the serine-protease inhibitor ecotin is able to inhibit a broad range of serine proteases that display a wide range of specificities. This specificity is driven by special structural features which allow unique flexibility upon binding to targets. Although frequently observed in many human/animal-associated bacteria, ecotin homologs may also be found in plant-associated taxa and environmental species. The purpose of this review is to provide an update on the biological importance, role in host–microbe interactions, and evolutionary relationship between ecotin orthologs isolated from Eukaryotic and Prokaryotic species across the Tree of Life.

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“…Essa proteína inibe fortemente as proteases do hospedeiro mamífero, como tripsina, quimotripsina, elastase de neutrófilos (NE) e fatores de coagulação do plasma (Garcia et al 2020).…”

Section: Confusumunclassified

Análise genômica comparativa e evolutiva do genoma do monoxeno Blechomonas sp. TCC303E

Tantaleán¹

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Acima de tudo, agradeço ao meu Deus, por seu mistério infinito onde dia a dia nós cientistas trabalhamos para entendê-lo um pouco mais.A minha mãe Lila Tantalean Campos e minha irmã Leidy Yannina Calderon Tantalean por serem minha força e empolgação para ser melhor a cada dia e a esse amor infinito que nos une, e também agradeço a meu amor de pandemia (agora noiva) Samara Spagnolo Sousa por sua parceria e companhia que me faz sentir bem cada dia. Agradeço também ao meu melhor amigo Hachi, por todos os momentos de diversão e tristeza juntos e aquele amor incondicional que os cães têm por seus donos, descanse em paz meu amigo.Ao Prof. João Marcelo Pereira Alves, mais conhecido pelo "J" por abrir as portas de seu laboratório para mim, pela oportunidade de fazer parte do time Labo 40; pela paciência e a confiança depositada para o desenvolvimento deste trabalho.Ao pessoal do lab. 40, Percy Omar Túllume Vergara e Anderson Carvalho Morais, obrigado pelo apoio e das dicas neste trabalho de mestre e também pela sua amizade. A Guillermo Uceda Campos pelas conversas e apoio sobre diversos assuntos.À Universidade de São Paulo, às agências de fomento à pesquisa CAPES pelo apoio financeiro. E para você, que embora seu nome não esteja listado nestes agradecimentos, mais que também contribui de alguma forma ou outra no desenvolvimento deste projeto. Só tenho a agradecer a todos vocês, muito obrigado! RESUMO O conhecimento dos tripanossomatídeos vem principalmente dos heteroxenos que foram muito mais estudados por terem impactos negativos na sociedade, deixando o maior grupo de tripanossomatídeos (monoxenos) quase esquecido por muito tempo. No entanto, os monoxenos se tornaram relevantes recentemente por serem bons modelos para entender a origem do parasitismo e a evolução dos heteroxenos, devido a serem basais a todos os heteroxenos importantes e compartilharem características genéticas comuns, além de co-infectar os hospedeiros desses heteroxenos. Diante dessa necessidade, este estudo tem como objetivo caracterizar o genoma do monoxeno Blechomonas sp. TCC303E (Classe Kinetoplastea; subfamília Blechomonadinae) através de técnicas bioinformáticas para analisar aspectos genômicos e evolutivos desse organismo, expandindo assim a disponibilidade de dados genéticos deste gênero, que até agora tinha disponível apenas a sequência genômica da espécie-tipo, Blechomonas ayalai. Blechomomas são parasitas extremamente polimórficos e exclusivos das pulgas (Siphonaptera), as quais podem transmitir agentes etiológicos de doenças veterinárias e médicas. Nossos resultados mostram uma montagem com uma cobertura média de 38,6 vezes, de boa qualidade e composta de 728 scaffolds, com N50 de ~61 kbp e uma média de GC de 50,3%. Um dos scaffolds contém DNA mitocondrial, com uma média de GC de 19,7%. Foram preditos 8.044 CDS, das quais foram anotadas funcionalmente 7.002, além de genes de RNAs: 71 tRNAs, 4 rRNAs, 62 RNAs não codificantes. A análise de sintenia mostrou que Blechomonas mantém uma arquitetura genômica semelhante à dos demais tripanossomat...

show abstract

Functional and structural characterization of an ecotin-like serine protease inhibitor from Trypanosoma cruzi

Cited by 6 publications

References 45 publications

Characterization of ecotin homologs from Campylobacter rectus and Campylobacter showae

Characterization of ecotin homologs from Campylobacter rectus and Campylobacter showae

Ecotin: A versatile protease inhibitor of bacteria and eukaryotes

Análise genômica comparativa e evolutiva do genoma do monoxeno Blechomonas sp. TCC303E

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