Open Access Research article

Anopheles salivary gland proteomes from major malaria vectors

Albin Fontaine1, Thierry Fusaï1, Sébastien Briolant1, Sylvain Buffet2, Claude Villard3, Emilie Baudelet4, Mathieu Pophillat5, Samuel Granjeaud6, Christophe Rogier17 and Lionel Almeras12*

Author Affiliations

1 Unité de Parasitologie – UMR6236, URMITE – IFR48, Antenne Marseille de l’Institut de Recherche Biomédicale des Armées (IRBA), BP 60109, Marseille Cedex 07, 13 262, France

2 Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes (URMITE), UMR 6236, Faculté de médecine, 27 Bd Jean Moulin, Marseille cedex 5, 13385, France

3 Plateau Protéomique Timone, FRE CNRS 2737 CISMET, université Aix-Marseille II, 27 Bd Jean Moulin, Marseille cedex 5, 13385, France

4 Plateforme de Spectrométrie de Masse et de Protéomique, Centre de Recherche en Cancérologie de Marseille, U1068, INSERM/Institut Paoli-Calmettes, 27 Bd Leï Roure, Marseille Cedex 9, BP 30059, 13273, France

5 Centre d'Immunologie de Marseille Luminy (CIML), Institut National de la Santé et de la Recherche Médicale, Centre National de la Recherche Scientifique, Université de la Méditerranée, Parc Scientifique de Luminy, Case 906, Marseille Cedex 9, Marseille, 13288, France

6 TAGC INSERM ERM 206, Parc Scientifique de Luminy, Case 928, Marseille Cedex 9, 13288, France

7 Institut Pasteur de Madagascar, B.P. 1274, Antananarivo, 101, Madagascar

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BMC Genomics 2012, 13:614  doi:10.1186/1471-2164-13-614

Published: 13 November 2012

Additional files

Additional file 1:

Average percentage identity between local alignments of secreted salivary proteins from six Anophelesspecies. All secreted salivary protein sequences from each Anopheles species (An. gambiae, An. arabiensis, An. stephensi, An. funestus, An. albimanus and An. darlingi) matching at least one other salivary protein in another species at 40% identity threshold (q.v., Additional file 2) were recovered and blasted again each other. The percentage identity of the best match (lowest E-value) was recovered with the protein NCBInr accession number and normalized BLAST scores were calculated based on raw BLAST scores and raw self-BLAST scores. When a unique salivary protein from target species B matched several proteins in reference species A, only the best match (lowest E-value) was selected. Average normalized BLAST scores ± SD and percentage identities are indicated in bold and summarized on Figure 1C.

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Additional file 2:

Hierarchical clustering of secreted salivary gland proteins from six Anophelesspecies. A three step clustering was performed at ≥ 90%, ≥ 70% and ≥ 40% identity threshold with the H-CD-HIT server on secreted salivary proteins from An. gambiae, An. arabiensis, An. stephensi, An. funestus, An. albimanus and An. darlingi. Clusters are sorted into protein families. The NCBI accession number is indicated for each protein. * indicate the representative (i.e., longest) protein sequence of each cluster. The percentage identity between the representative protein sequence (*) and other protein sequences is given for each cluster. Protein in bold are new clusterised proteins at each identity threshold. Results from this table are graphically represented on Figure 2.

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Additional file 3:

Proteins identified by MS in salivary gland extracts of four Anophelesspecies. All MS/MS spectra resulting to every protein bands from each species (An. gambiae, An. arabiensis, An. stephensi and An. albimanus) were gathered and searched on sequence databases of the four Anopheles species together. A list of all unique proteins identified in salivary gland extracts in both replicates is presented for each Anopheles species. Salivary gland proteins were sorted according to their signal peptide prediction (SignalP Neural Network) [63,65] to discriminate secreted proteins from housekeeping ones.

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Additional file 4:

Hierarchical clustering of putative secreted proteins identified in Anophelessalivary gland extracts. Proteins from An. gambiae, An. arabiensis, An. stephensi and An. albimanus SGEs were identified by mass spectrometry after in-gel trypsin digestion. Protein sequences were submitted to SignalP 3.0 server [65] to select putative secreted proteins and were hierarchically clustered at ≥ 90%, ≥ 70% and ≥ 40% identity threshold with CD-HIT web server [68]. * indicate the representative (i.e., longest) protein sequence of each cluster. Anopheles species in which secreted salivary proteins were identified are indicated. The last common taxon encompassing homologous proteins at the genus level is indicated according to in silico results (q.v. Additional file 2). n.a.: non-available (i.e., uncharacterized protein sequences that were not recovered in the in silico analysis). Lines in bold indicate proteins identified in antigenic bands (Figure 5A, Additional file 5). AGA, An. gambiae; AGA#, An. gambiae PEST strain (Pink Eye STandard); AAR, An. arabiensis; AST, An. stephensi; AAL, An. albimanus; MW: Molecular weight.

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Additional file 5:

Proteins identified by MS in antigenic protein bands from salivary gland extracts from Anophelesspecies. Salivary gland proteins identified in each antigenic protein band from An. gambiae, An. arabiensis, An. stephensi and An. albimanus SGEs are indicated. Band numbers correspond to those indicated on Figure 5C.

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Additional file 6:

Alignment of members of the GE-rich/30 kDa/anti-platelet protein family from An. gambiae, An. stephensi and An. albimanus. The numbers in the sequence titles indicate the NCBI accession number.

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