[Comparison of three protein extraction methods for proteomic analysis of Alexandrium tamarense with two-dimensional
Yawei Yuan1, Wei Zheng, Shaojing Li
1State Key Laboratory for Marine Environmental Science, Xiamen University, Xiamen 361005, China. yawei1983@126.com
Objective:
In order to find the best extraction method for proteins of Alexandrium tamarense for two-dimensional electrophoresis (2-DE) analysis.
Methods:
Three methods for extracting proteins from A. tamarense were compared, including trihydroxymethyl aminomethane (Tris-HCl ) buffer extraction, trichloroacetic acid (TCA)/acetone precipitation and lysis buffer extraction. Alga was cultivated in normal f/2 media (control) and supplemented with algicidal substances. Proteins obtained using the best extraction method were separated with 2-DE. Protein-expression differences were identified using matrix-assisted laser desorption/ionization-time of flight ( MALDI-TOF) mass spectrometry (MS).
Results:
Among the three protein extraction methods, lysis buffer extraction shows the best detection of the number and quality of protein spots with a clear background. Then, the lysis buffer extraction method was successfully applied to profiling protein expression in algicidal substances stress conditions and 14 differential expression proteins were identified using MALDI-TOF/MS.
Conclusion:
Lysis buffer extraction was the most effective protein extraction method for Alexandrium tamarense.
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