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An insight into the sialome of Simulium guianense (DIPTERA:SIMulIIDAE), the main vector of River Blindness Disease in Brazil.

Chagas AC, Calvo E, Pimenta PF, Ribeiro JM - BMC Genomics (2011)

Bottom Line: Insect-specific families were also found.About 63.4% of all secreted products revealed protein families found only in Simulium.Additionally, we found a novel peptide similar to kunitoxin with a structure distantly related to serine protease inhibitors.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, 12735 Twinbrook Parkway, National Institutes of Health, Rockville, Maryland 20892-8132, USA.

ABSTRACT

Background: Little is known about the composition and function of the saliva in black flies such as Simulium guianense, the main vector of river blindness disease in Brazil. The complex salivary potion of hematophagous arthropods counteracts their host's hemostasis, inflammation, and immunity.

Results: Transcriptome analysis revealed ubiquitous salivary protein families--such as the Antigen-5, Yellow, Kunitz domain, and serine proteases--in the S. guianense sialotranscriptome. Insect-specific families were also found. About 63.4% of all secreted products revealed protein families found only in Simulium. Additionally, we found a novel peptide similar to kunitoxin with a structure distantly related to serine protease inhibitors. This study revealed a relative increase of transcripts of the SVEP protein family when compared with Simulium vittatum and S. nigrimanum sialotranscriptomes. We were able to extract coding sequences from 164 proteins associated with blood and sugar feeding, the majority of which were confirmed by proteome analysis.

Conclusions: Our results contribute to understanding the role of Simulium saliva in transmission of Onchocerca volvulus and evolution of salivary proteins in black flies. It also consists of a platform for mining novel anti-hemostatic compounds, vaccine candidates against filariasis, and immuno-epidemiologic markers of vector exposure.

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Related in: MedlinePlus

1-D gel electrophoresis of Simulium guianense salivary gland homogenates. The numbers at the left indicate the mol wt of the protein standards (kDa), shown in the left lane. The right gel lane shows the separation of the salivary gland proteins. The grid at the right (F1-32) represents the gel slices submitted for tryptic digest and tandem mass spectrometry identification.
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Figure 2: 1-D gel electrophoresis of Simulium guianense salivary gland homogenates. The numbers at the left indicate the mol wt of the protein standards (kDa), shown in the left lane. The right gel lane shows the separation of the salivary gland proteins. The grid at the right (F1-32) represents the gel slices submitted for tryptic digest and tandem mass spectrometry identification.

Mentions: Several clusters of sequences coding for H and S polypeptides indicated in Additional File 1 are abundant and complete enough to extract novel consensus sequences. A total of 174 novel sequences--164 of which code for S proteins-- are grouped together in Additional File 2. With this database, we characterized the proteome via analysis of SDS-PAGE separated proteins that were tryptic digested and submitted to MS/MS analysis (Figure 2). The results of this experiment are integrated within the description of the deduced proteins from the transcriptome analysis. Here, we used proteome analysis to confirm 28 of the 32 protein families found in this sialotranscriptome (Additional File 2), which are described in more detail below. The reader is here informed that the introduction of the diverse protein families may contain text previously used in our publications on Simulium sialomes [14,15] and such text will appear in "quotes".


An insight into the sialome of Simulium guianense (DIPTERA:SIMulIIDAE), the main vector of River Blindness Disease in Brazil.

Chagas AC, Calvo E, Pimenta PF, Ribeiro JM - BMC Genomics (2011)

1-D gel electrophoresis of Simulium guianense salivary gland homogenates. The numbers at the left indicate the mol wt of the protein standards (kDa), shown in the left lane. The right gel lane shows the separation of the salivary gland proteins. The grid at the right (F1-32) represents the gel slices submitted for tryptic digest and tandem mass spectrometry identification.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3285218&req=5

Figure 2: 1-D gel electrophoresis of Simulium guianense salivary gland homogenates. The numbers at the left indicate the mol wt of the protein standards (kDa), shown in the left lane. The right gel lane shows the separation of the salivary gland proteins. The grid at the right (F1-32) represents the gel slices submitted for tryptic digest and tandem mass spectrometry identification.
Mentions: Several clusters of sequences coding for H and S polypeptides indicated in Additional File 1 are abundant and complete enough to extract novel consensus sequences. A total of 174 novel sequences--164 of which code for S proteins-- are grouped together in Additional File 2. With this database, we characterized the proteome via analysis of SDS-PAGE separated proteins that were tryptic digested and submitted to MS/MS analysis (Figure 2). The results of this experiment are integrated within the description of the deduced proteins from the transcriptome analysis. Here, we used proteome analysis to confirm 28 of the 32 protein families found in this sialotranscriptome (Additional File 2), which are described in more detail below. The reader is here informed that the introduction of the diverse protein families may contain text previously used in our publications on Simulium sialomes [14,15] and such text will appear in "quotes".

Bottom Line: Insect-specific families were also found.About 63.4% of all secreted products revealed protein families found only in Simulium.Additionally, we found a novel peptide similar to kunitoxin with a structure distantly related to serine protease inhibitors.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, 12735 Twinbrook Parkway, National Institutes of Health, Rockville, Maryland 20892-8132, USA.

ABSTRACT

Background: Little is known about the composition and function of the saliva in black flies such as Simulium guianense, the main vector of river blindness disease in Brazil. The complex salivary potion of hematophagous arthropods counteracts their host's hemostasis, inflammation, and immunity.

Results: Transcriptome analysis revealed ubiquitous salivary protein families--such as the Antigen-5, Yellow, Kunitz domain, and serine proteases--in the S. guianense sialotranscriptome. Insect-specific families were also found. About 63.4% of all secreted products revealed protein families found only in Simulium. Additionally, we found a novel peptide similar to kunitoxin with a structure distantly related to serine protease inhibitors. This study revealed a relative increase of transcripts of the SVEP protein family when compared with Simulium vittatum and S. nigrimanum sialotranscriptomes. We were able to extract coding sequences from 164 proteins associated with blood and sugar feeding, the majority of which were confirmed by proteome analysis.

Conclusions: Our results contribute to understanding the role of Simulium saliva in transmission of Onchocerca volvulus and evolution of salivary proteins in black flies. It also consists of a platform for mining novel anti-hemostatic compounds, vaccine candidates against filariasis, and immuno-epidemiologic markers of vector exposure.

Show MeSH
Related in: MedlinePlus