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Analysis of the Epithelial Damage Produced by Entamoeba histolytica Infection
Published on: June 12, 2014
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Analysis of Entamoeba histolytica Membrane Proteome Using Three Extraction Methods
Jorim Ujang1, Asmahani Azira Abdu Sani2, Boon Huat Lim3
1Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800, Penang, Malaysia.
Proteomics
|October 5, 2018
Summary
This study evaluated three methods for extracting Entamoeba histolytica membrane proteins for mass spectrometry. The conventional method and ProteoExtract yielded more proteins, while ProteoPrep offered higher membrane protein selectivity.
Area of Science:
- Proteomics
- Parasitology
- Molecular Biology
Background:
- Entamoeba histolytica causes amoebiasis, a significant human parasitic disease.
- Membrane proteins of E. histolytica are crucial for pathogenesis but remain largely uncharacterized.
- Efficient protein extraction is essential for comprehensive proteomic analysis.
Purpose of the Study:
- To evaluate and compare three protein extraction methods for studying the E. histolytica membrane proteome.
- To identify and characterize E. histolytica membrane proteins using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS).
- To assess the yield and selectivity of membrane proteins obtained from different extraction techniques.
Main Methods:
- Comparison of three protein extraction methods: ProteoExtract (Calbiochem), ProteoPrep (Sigma), and a conventional laboratory method.
- Analysis of extracted proteins using Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry (LC-ESI-MS/MS).
- In silico prediction and analysis of membrane protein fractions.
Main Results:
- ProteoExtract and the conventional method yielded higher total protein amounts compared to ProteoPrep.
- LC-ESI-MS/MS identified 456, 482, and 551 membrane fraction proteins using ProteoExtract, ProteoPrep, and the conventional method, respectively.
- ProteoPrep demonstrated the highest selectivity for membrane proteins, with 45% predicted membrane proteins, while ProteoExtract and the conventional method yielded 21% and 34%, respectively.
Conclusions:
- A total of 828 E. histolytica membrane fraction proteins were identified, including 249 predicted membrane proteins.
- The choice of extraction method impacts both protein yield and the selectivity of membrane protein identification.
- ProteoPrep is recommended for studies prioritizing membrane protein selectivity, while ProteoExtract and conventional methods may be preferred for higher overall protein yield.
- The study provides valuable proteomic data for E. histolytica, available via ProteomeXchange (PXD010171).
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