|Proteome analysis of Cry4Ba toxin-interacting Aedes aegypti lipid rafts using geLC-MS/MS.|
|PMID: 23153095 Owner: NLM Status: MEDLINE|
|Lipid rafts are microdomains in the plasma membrane of eukaryotic cells. Among their many functions, lipid rafts are involved in cell toxicity caused by pore forming bacterial toxins including Bacillus thuringiensis (Bt) Cry toxins. We isolated lipid rafts from brush border membrane vesicles (BBMV) of Aedes aegypti larvae as a detergent resistant membrane (DRM) fraction on density gradients. Cholesterol, aminopeptidase (APN), alkaline phosphatase (ALP) and the raft marker flotillin were preferentially partitioned into the lipid raft fraction. When mosquitocidal Cry4Ba toxin was preincubated with BBMV, Cry4Ba localized to lipid rafts. A proteomic approach based on one-dimensional gel electrophoresis, in-gel trypsin digestion, followed by liquid chromatography-mass spectrometry (geLC-MS/MS) identified a total of 386 proteins. Of which many are typical lipid raft marker proteins including flotillins and glycosylphosphatidylinositol (GPI)-anchored proteins. Identified raft proteins were annotated in silico for functional and physicochemical characteristics. Parameters such as distribution of isoelectric point, molecular mass, and predicted post-translational modifications relevant to lipid raft proteins (GPI anchorage and myristoylation or palmitoylation) were analyzed for identified proteins in the DRM fraction. From a functional point of view, this study identified proteins implicated in Cry toxin interactions as well as membrane-associated proteins expressed in the mosquito midgut that have potential relevance to mosquito biology and vector management.|
|Krishnareddy Bayyareddy; Xiang Zhu; Ron Orlando; Michael J Adang|
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|Type: Journal Article; Research Support, N.I.H., Extramural Date: 2012-11-27|
|Title: Journal of proteome research Volume: 11 ISSN: 1535-3907 ISO Abbreviation: J. Proteome Res. Publication Date: 2012 Dec|
|Created Date: 2012-12-11 Completed Date: 2013-05-23 Revised Date: 2013-12-12|
Medline Journal Info:
|Nlm Unique ID: 101128775 Medline TA: J Proteome Res Country: United States|
|Languages: eng Pagination: 5843-55 Citation Subset: IM|
|APA/MLA Format Download EndNote Download BibTex|
Alkaline Phosphatase / chemistry
Aminopeptidases / chemistry
Bacterial Proteins / chemistry*
Chromatography, Liquid / methods*
Electrophoresis, Polyacrylamide Gel
Endotoxins / chemistry*
Escherichia coli / chemistry
Glycosylphosphatidylinositols / chemistry
Hemolysin Proteins / chemistry*
Larva / chemistry, enzymology
Membrane Microdomains / chemistry*
Membrane Proteins / chemistry
Microvilli / chemistry
Protein Interaction Mapping
Protein Processing, Post-Translational
Proteome / analysis*, chemistry
Proteomics / methods
Receptors, Cell Surface / chemistry
Tandem Mass Spectrometry / methods
|R01 AI 29092/AI/NIAID NIH HHS; R01 AI029092/AI/NIAID NIH HHS|
|0/Bacterial Proteins; 0/Detergents; 0/Endotoxins; 0/Glycosylphosphatidylinositols; 0/Hemolysin Proteins; 0/Membrane Proteins; 0/Proteome; 0/Receptors, Cell Surface; 0/flotillins; 0/insecticidal crystal protein, Bacillus Thuringiensis; 9002-93-1/Octoxynol; EC 22.214.171.124/Alkaline Phosphatase; EC 3.4.11.-/Aminopeptidases|
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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