Collection and analysis of salivary proteins from the biting midge Culicoides nubeculosus (Diptera: Ceratopogonidae).

2.50
Hdl Handle:
http://hdl.handle.net/10033/30455
Title:
Collection and analysis of salivary proteins from the biting midge Culicoides nubeculosus (Diptera: Ceratopogonidae).
Authors:
Langner, Kathrin F A; Darpel, Karin E; Denison, Eric; Drolet, Barbara S; Leibold, Wolfgang; Mellor, Philip S; Mertens, Peter P C; Nimtz, Manfred; Greiser-Wilke, Irene
Abstract:
Salivary proteins of hematophagous Culicoides spp. are thought to play an important role in pathogen transmission and skin hypersensitivity. Analysis of these proteins, however, has been problematic due to the difficulty in obtaining adequate amounts of secreted Culicoides saliva. In the current study, a collection method for midge saliva was developed. Over a 3-d period, 3- to 5-d-old male and female Culicoides nubeculosus Meigen (Diptera: Ceratopogonidae) were repeatedly placed onto the collection system and allowed to deposit saliva into a filter. Salivary products were eluted from the filters and evaluated by gel electrophoresis and mass spectrometry as well as by intradermal testing and determination of clotting time. Gel electrophoresis revealed approximately 55 protein spots displaying relative molecular masses from 5 to 67 kDa and isoelectric points ranging from 4.5 to 9.8. The majority of molecular species analyzed by mass spectrometry showed high convergence with salivary proteins recently obtained from a cDNA library of Culicoides sonorensis Wirth & Jones, including proteins involved in sugarmeal digestion, defense, and coagulation inhibition as well as members of the D7 family and unclassified salivary proteins. In addition, the proteome analysis revealed a number of peptides that were related to proteins from insect species other than Culicoides. Intradermal injection of the saliva in human skin produced edema, vasodilatation, and pruritus. The anticoagulant activity of the saliva was demonstrated by significantly prolonged clotting times for human platelets. The potential role of the identified salivary proteins in the transmission of pathogens and the induction of allergies is discussed.
Affiliation:
USDA-ARS, Arthropod-Borne Animal Diseases Research Laboratory, 1000 E. University Ave., Laramie, WY 82071, USA. klangner@uwyo.edu
Citation:
Collection and analysis of salivary proteins from the biting midge Culicoides nubeculosus (Diptera: Ceratopogonidae). 2007, 44 (2):238-48 J. Med. Entomol.
Journal:
Journal of medical entomology
Issue Date:
Mar-2007
URI:
http://hdl.handle.net/10033/30455
PubMed ID:
17427692
Type:
Article
Language:
en
ISSN:
0022-2585
Appears in Collections:
publications of the research group cellular proteom research (CPRO)

Full metadata record

DC FieldValue Language
dc.contributor.authorLangner, Kathrin F A-
dc.contributor.authorDarpel, Karin E-
dc.contributor.authorDenison, Eric-
dc.contributor.authorDrolet, Barbara S-
dc.contributor.authorLeibold, Wolfgang-
dc.contributor.authorMellor, Philip S-
dc.contributor.authorMertens, Peter P C-
dc.contributor.authorNimtz, Manfred-
dc.contributor.authorGreiser-Wilke, Irene-
dc.date.accessioned2008-06-25T10:45:06Z-
dc.date.available2008-06-25T10:45:06Z-
dc.date.issued2007-03-
dc.identifier.citationCollection and analysis of salivary proteins from the biting midge Culicoides nubeculosus (Diptera: Ceratopogonidae). 2007, 44 (2):238-48 J. Med. Entomol.en
dc.identifier.issn0022-2585-
dc.identifier.pmid17427692-
dc.identifier.urihttp://hdl.handle.net/10033/30455-
dc.description.abstractSalivary proteins of hematophagous Culicoides spp. are thought to play an important role in pathogen transmission and skin hypersensitivity. Analysis of these proteins, however, has been problematic due to the difficulty in obtaining adequate amounts of secreted Culicoides saliva. In the current study, a collection method for midge saliva was developed. Over a 3-d period, 3- to 5-d-old male and female Culicoides nubeculosus Meigen (Diptera: Ceratopogonidae) were repeatedly placed onto the collection system and allowed to deposit saliva into a filter. Salivary products were eluted from the filters and evaluated by gel electrophoresis and mass spectrometry as well as by intradermal testing and determination of clotting time. Gel electrophoresis revealed approximately 55 protein spots displaying relative molecular masses from 5 to 67 kDa and isoelectric points ranging from 4.5 to 9.8. The majority of molecular species analyzed by mass spectrometry showed high convergence with salivary proteins recently obtained from a cDNA library of Culicoides sonorensis Wirth & Jones, including proteins involved in sugarmeal digestion, defense, and coagulation inhibition as well as members of the D7 family and unclassified salivary proteins. In addition, the proteome analysis revealed a number of peptides that were related to proteins from insect species other than Culicoides. Intradermal injection of the saliva in human skin produced edema, vasodilatation, and pruritus. The anticoagulant activity of the saliva was demonstrated by significantly prolonged clotting times for human platelets. The potential role of the identified salivary proteins in the transmission of pathogens and the induction of allergies is discussed.en
dc.language.isoenen
dc.subject.meshAnimalsen
dc.subject.meshAnticoagulantsen
dc.subject.meshCeratopogonidaeen
dc.subject.meshElectrophoresis, Gel, Two-Dimensionalen
dc.subject.meshFemaleen
dc.subject.meshHumansen
dc.subject.meshInjections, Intradermalen
dc.subject.meshInsect Vectorsen
dc.subject.meshIntradermal Testsen
dc.subject.meshMaleen
dc.subject.meshMass Spectrometryen
dc.subject.meshSalivaen
dc.subject.meshSalivary Proteinsen
dc.subject.meshVasodilator Agentsen
dc.titleCollection and analysis of salivary proteins from the biting midge Culicoides nubeculosus (Diptera: Ceratopogonidae).en
dc.typeArticleen
dc.contributor.departmentUSDA-ARS, Arthropod-Borne Animal Diseases Research Laboratory, 1000 E. University Ave., Laramie, WY 82071, USA. klangner@uwyo.eduen
dc.identifier.journalJournal of medical entomologyen

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