Susceptibility of Phytomonas serpens to calpain inhibitors in vitro: interference on the proliferation, ultrastructure, cysteine peptidase expression and interaction with the invertebrate host
A pleiotropic response to the calpain inhibitor MDL28170 was detected in the tomato parasite Phytomonas serpens. Ultrastructural studies revealed that MDL28170 caused mitochondrial swelling, shortening of flagellum and disruption of trans Golgi network. This effect was correlated to the inhibition i...
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| Format: | Article |
| Language: | English |
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Fundação Oswaldo Cruz (FIOCRUZ)
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| Series: | Memorias do Instituto Oswaldo Cruz |
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| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762016005026103&lng=en&tlng=en |
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| author | Simone Santiago Carvalho de Oliveira Diego de Souza Gonçalves Aline dos Santos Garcia-Gomes Inês Correa Gonçalves Sergio Henrique Seabra Rubem Figueiredo Menna-Barreto Angela Hampshire de Carvalho Santos Lopes Claudia Masini D’Avila-Levy André Luis Souza dos Santos Marta Helena Branquinha |
| author_facet | Simone Santiago Carvalho de Oliveira Diego de Souza Gonçalves Aline dos Santos Garcia-Gomes Inês Correa Gonçalves Sergio Henrique Seabra Rubem Figueiredo Menna-Barreto Angela Hampshire de Carvalho Santos Lopes Claudia Masini D’Avila-Levy André Luis Souza dos Santos Marta Helena Branquinha |
| author_sort | Simone Santiago Carvalho de Oliveira |
| collection | DOAJ |
| description | A pleiotropic response to the calpain inhibitor MDL28170 was detected in the tomato parasite Phytomonas serpens. Ultrastructural studies revealed that MDL28170 caused mitochondrial swelling, shortening of flagellum and disruption of trans Golgi network. This effect was correlated to the inhibition in processing of cruzipain-like molecules, which presented an increase in expression paralleled by decreased proteolytic activity. Concomitantly, a calcium-dependent cysteine peptidase was detected in the parasite extract, the activity of which was repressed by pre-incubation of parasites with MDL28170. Flow cytometry and Western blotting analyses revealed the differential expression of calpain-like proteins (CALPs) in response to the pre-incubation of parasites with the MDL28170, and confocal fluorescence microscopy confirmed their surface location. The interaction of promastigotes with explanted salivary glands of the insect Oncopeltus fasciatus was reduced when parasites were pre-treated with MDL28170, which was correlated to reduced levels of surface cruzipain-like and gp63-like molecules. Treatment of parasites with anti-Drosophila melanogaster (Dm) calpain antibody also decreased the adhesion process. Additionally, parasites recovered from the interaction process presented higher levels of surface cruzipain-like and gp63-like molecules, with similar levels of CALPs cross-reactive to anti-Dm-calpain antibody. The results confirm the importance of exploring the use of calpain inhibitors in studying parasites’ physiology. |
| first_indexed | 2024-03-12T18:22:39Z |
| format | Article |
| id | doaj.art-f5964f2b01814c768b5f580cf58d538d |
| institution | Directory Open Access Journal |
| issn | 1678-8060 |
| language | English |
| last_indexed | 2024-03-12T18:22:39Z |
| publisher | Fundação Oswaldo Cruz (FIOCRUZ) |
| record_format | Article |
| series | Memorias do Instituto Oswaldo Cruz |
| spelling | doaj.art-f5964f2b01814c768b5f580cf58d538d2023-08-02T08:46:26ZengFundação Oswaldo Cruz (FIOCRUZ)Memorias do Instituto Oswaldo Cruz1678-8060010.1590/0074-02760160270S0074-02762016005026103Susceptibility of Phytomonas serpens to calpain inhibitors in vitro: interference on the proliferation, ultrastructure, cysteine peptidase expression and interaction with the invertebrate hostSimone Santiago Carvalho de OliveiraDiego de Souza GonçalvesAline dos Santos Garcia-GomesInês Correa GonçalvesSergio Henrique SeabraRubem Figueiredo Menna-BarretoAngela Hampshire de Carvalho Santos LopesClaudia Masini D’Avila-LevyAndré Luis Souza dos SantosMarta Helena BranquinhaA pleiotropic response to the calpain inhibitor MDL28170 was detected in the tomato parasite Phytomonas serpens. Ultrastructural studies revealed that MDL28170 caused mitochondrial swelling, shortening of flagellum and disruption of trans Golgi network. This effect was correlated to the inhibition in processing of cruzipain-like molecules, which presented an increase in expression paralleled by decreased proteolytic activity. Concomitantly, a calcium-dependent cysteine peptidase was detected in the parasite extract, the activity of which was repressed by pre-incubation of parasites with MDL28170. Flow cytometry and Western blotting analyses revealed the differential expression of calpain-like proteins (CALPs) in response to the pre-incubation of parasites with the MDL28170, and confocal fluorescence microscopy confirmed their surface location. The interaction of promastigotes with explanted salivary glands of the insect Oncopeltus fasciatus was reduced when parasites were pre-treated with MDL28170, which was correlated to reduced levels of surface cruzipain-like and gp63-like molecules. Treatment of parasites with anti-Drosophila melanogaster (Dm) calpain antibody also decreased the adhesion process. Additionally, parasites recovered from the interaction process presented higher levels of surface cruzipain-like and gp63-like molecules, with similar levels of CALPs cross-reactive to anti-Dm-calpain antibody. The results confirm the importance of exploring the use of calpain inhibitors in studying parasites’ physiology.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762016005026103&lng=en&tlng=enPhytomonascalpain-like proteinscysteine peptidasecruzipainGp63Oncopeltus fasciatus |
| spellingShingle | Simone Santiago Carvalho de Oliveira Diego de Souza Gonçalves Aline dos Santos Garcia-Gomes Inês Correa Gonçalves Sergio Henrique Seabra Rubem Figueiredo Menna-Barreto Angela Hampshire de Carvalho Santos Lopes Claudia Masini D’Avila-Levy André Luis Souza dos Santos Marta Helena Branquinha Susceptibility of Phytomonas serpens to calpain inhibitors in vitro: interference on the proliferation, ultrastructure, cysteine peptidase expression and interaction with the invertebrate host Memorias do Instituto Oswaldo Cruz Phytomonas calpain-like proteins cysteine peptidase cruzipain Gp63 Oncopeltus fasciatus |
| title | Susceptibility of Phytomonas serpens to calpain inhibitors in vitro: interference on the proliferation, ultrastructure, cysteine peptidase expression and interaction with the invertebrate host |
| title_full | Susceptibility of Phytomonas serpens to calpain inhibitors in vitro: interference on the proliferation, ultrastructure, cysteine peptidase expression and interaction with the invertebrate host |
| title_fullStr | Susceptibility of Phytomonas serpens to calpain inhibitors in vitro: interference on the proliferation, ultrastructure, cysteine peptidase expression and interaction with the invertebrate host |
| title_full_unstemmed | Susceptibility of Phytomonas serpens to calpain inhibitors in vitro: interference on the proliferation, ultrastructure, cysteine peptidase expression and interaction with the invertebrate host |
| title_short | Susceptibility of Phytomonas serpens to calpain inhibitors in vitro: interference on the proliferation, ultrastructure, cysteine peptidase expression and interaction with the invertebrate host |
| title_sort | susceptibility of phytomonas serpens to calpain inhibitors in vitro interference on the proliferation ultrastructure cysteine peptidase expression and interaction with the invertebrate host |
| topic | Phytomonas calpain-like proteins cysteine peptidase cruzipain Gp63 Oncopeltus fasciatus |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762016005026103&lng=en&tlng=en |
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