Expression of active trypsin-like serine peptidases in the midgut of sugar-feeding female Anopheles aquasalis
Abstract Background Anopheles aquasalis is a dipteran of the family Culicidae that is widely distributed in the coastal regions of South and Central America. This species acts as a vector of Plasmodium vivax, an important etiological agent of malaria, which represents a serious public health problem...
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| Format: | Article |
| Language: | English |
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BMC
2015-05-01
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| Series: | Parasites & Vectors |
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| Online Access: | https://doi.org/10.1186/s13071-015-0908-0 |
| _version_ | 1797811818549215232 |
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| author | Geovane Dias-Lopes Andre Borges-Veloso Leonardo Saboia-Vahia Gilberto B. Domont Constança Britto Patricia Cuervo Jose Batista De Jesus |
| author_facet | Geovane Dias-Lopes Andre Borges-Veloso Leonardo Saboia-Vahia Gilberto B. Domont Constança Britto Patricia Cuervo Jose Batista De Jesus |
| author_sort | Geovane Dias-Lopes |
| collection | DOAJ |
| description | Abstract Background Anopheles aquasalis is a dipteran of the family Culicidae that is widely distributed in the coastal regions of South and Central America. This species acts as a vector of Plasmodium vivax, an important etiological agent of malaria, which represents a serious public health problem. In mosquitoes, trypsin-like serine proteases are important in blood meal digestion, immune responses and reproductive functions. The study of peptidases expressed in the mosquito midgut is essential to understanding the mechanisms of parasite-host interaction and the physiological process of nutrient digestion. Methods Our study aimed to identify and characterize the proteolytic activities in the midgut of sugar-fed An. aquasalis females using zymographic analyses (substrate-SDS-PAGE), in-solution assays and mass spectrometry. Results Here, we used a zymographic analysis to further biochemically characterize the proteolytic profile of the midgut of sugar-feeding An. aquasalis females. The trypsin peptidases migrated between ~17 and ~76 kDa and displayed higher proteolytic activities between pH 7.5 and 10 and at temperatures between 37 °C and 50 °C. Four putative trypsin-like serine peptidases were identified using mass spectrometry and data mining. The molecular masses of these peptidases were similar to those observed using zymography, which suggested that these peptidases could be responsible for some of the observed proteolytic bands. Conclusions Taken together, our results contribute to the gene annotation of the unknown genome of this species, to the tissue location of these peptidases, and to the functional prediction of these crucial enzymes, which all impact further studies of this species. |
| first_indexed | 2024-03-13T07:29:27Z |
| format | Article |
| id | doaj.art-697d3a0a4c504f63a1a3478816787769 |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-03-13T07:29:27Z |
| publishDate | 2015-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Parasites & Vectors |
| spelling | doaj.art-697d3a0a4c504f63a1a34788167877692023-06-04T11:11:41ZengBMCParasites & Vectors1756-33052015-05-018111010.1186/s13071-015-0908-0Expression of active trypsin-like serine peptidases in the midgut of sugar-feeding female Anopheles aquasalisGeovane Dias-Lopes0Andre Borges-Veloso1Leonardo Saboia-Vahia2Gilberto B. Domont3Constança Britto4Patricia Cuervo5Jose Batista De Jesus6Laboratório de Biologia Molecular e Doenças Endêmicas – Instituto Oswaldo Cruz, FIOCRUZLaboratório de Biologia Molecular e Doenças Endêmicas – Instituto Oswaldo Cruz, FIOCRUZLaboratório de Pesquisa em Leishmaniose – Instituto Oswaldo Cruz, FIOCRUZLaboratório de Química de Proteínas, Departamento de Química, UFRJLaboratório de Biologia Molecular e Doenças Endêmicas – Instituto Oswaldo Cruz, FIOCRUZLaboratório de Pesquisa em Leishmaniose – Instituto Oswaldo Cruz, FIOCRUZLaboratório de Biologia Molecular e Doenças Endêmicas – Instituto Oswaldo Cruz, FIOCRUZAbstract Background Anopheles aquasalis is a dipteran of the family Culicidae that is widely distributed in the coastal regions of South and Central America. This species acts as a vector of Plasmodium vivax, an important etiological agent of malaria, which represents a serious public health problem. In mosquitoes, trypsin-like serine proteases are important in blood meal digestion, immune responses and reproductive functions. The study of peptidases expressed in the mosquito midgut is essential to understanding the mechanisms of parasite-host interaction and the physiological process of nutrient digestion. Methods Our study aimed to identify and characterize the proteolytic activities in the midgut of sugar-fed An. aquasalis females using zymographic analyses (substrate-SDS-PAGE), in-solution assays and mass spectrometry. Results Here, we used a zymographic analysis to further biochemically characterize the proteolytic profile of the midgut of sugar-feeding An. aquasalis females. The trypsin peptidases migrated between ~17 and ~76 kDa and displayed higher proteolytic activities between pH 7.5 and 10 and at temperatures between 37 °C and 50 °C. Four putative trypsin-like serine peptidases were identified using mass spectrometry and data mining. The molecular masses of these peptidases were similar to those observed using zymography, which suggested that these peptidases could be responsible for some of the observed proteolytic bands. Conclusions Taken together, our results contribute to the gene annotation of the unknown genome of this species, to the tissue location of these peptidases, and to the functional prediction of these crucial enzymes, which all impact further studies of this species.https://doi.org/10.1186/s13071-015-0908-0Anopheles aquasalisMidgutZymographyTrypsin-like serine peptidasesProteomicsMass spectrometry |
| spellingShingle | Geovane Dias-Lopes Andre Borges-Veloso Leonardo Saboia-Vahia Gilberto B. Domont Constança Britto Patricia Cuervo Jose Batista De Jesus Expression of active trypsin-like serine peptidases in the midgut of sugar-feeding female Anopheles aquasalis Parasites & Vectors Anopheles aquasalis Midgut Zymography Trypsin-like serine peptidases Proteomics Mass spectrometry |
| title | Expression of active trypsin-like serine peptidases in the midgut of sugar-feeding female Anopheles aquasalis |
| title_full | Expression of active trypsin-like serine peptidases in the midgut of sugar-feeding female Anopheles aquasalis |
| title_fullStr | Expression of active trypsin-like serine peptidases in the midgut of sugar-feeding female Anopheles aquasalis |
| title_full_unstemmed | Expression of active trypsin-like serine peptidases in the midgut of sugar-feeding female Anopheles aquasalis |
| title_short | Expression of active trypsin-like serine peptidases in the midgut of sugar-feeding female Anopheles aquasalis |
| title_sort | expression of active trypsin like serine peptidases in the midgut of sugar feeding female anopheles aquasalis |
| topic | Anopheles aquasalis Midgut Zymography Trypsin-like serine peptidases Proteomics Mass spectrometry |
| url | https://doi.org/10.1186/s13071-015-0908-0 |
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