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Open AccessArticle

Comparison of Five TRIzol-Based Protein Preparation Methods for 2-DE Production From Challenging Marine Dinoflagellate Samples: A Case Study on Two Benthic Prorocentrum Species

Department of Science, School of Science and Technology, The Open University of Hong Kong, Hong Kong, China
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J. Mar. Sci. Eng. 2020, 8(5), 363; https://doi.org/10.3390/jmse8050363
Received: 14 April 2020 / Revised: 9 May 2020 / Accepted: 18 May 2020 / Published: 21 May 2020
(This article belongs to the Special Issue Biocomplexity and Microbial Diversity in a Marine System)
Two-dimensional gel electrophoresis (2-DE) is a major element of conventional gel-based proteomics, which resolves complex protein mixtures. Protein extraction with the removal of interfering substances from the sample remains the key to producing high-quality 2-DE profiles. Marine dinoflagellates contain large endogenous amounts of salts, nucleic acids, polysaccharides, phenolic compounds, pigments, and other interfering compounds. These substances are detrimental to the quality of gel images. Protein preparation using TRIzol reagent is a promising method for producing high-quality 2-DE profiles for dinoflagellate samples. In addition to its remarkable performance, the TRIzol method’s several advantages have made it a popular and widely used method in the field of 2-DE sample preparation. Nonetheless, the quality of 2-DE of samples from certain dinoflagellate species is not as high as previously reported when the same TRIzol protocol is applied. Therefore, modifications to the original TRIzol method are required to remove interfering substances from those challenging dinoflagellate samples. In this study, the original TRIzol method and four modified methods, namely the aliquot TRIzol method, re-TRIzol method, TRIzol method with a commercial clean-up kit, and TRIzol method with trichloroacetic acid/acetone precipitation, were compared. Performance of these five methods in terms of protein yield, background signal, and resolution and number of protein spots was investigated on samples from two benthic Prorocentrum species: P. lima and P. hoffmannianum. Our results demonstrated that high-quality 2-DE could be achieved from P. lima samples prepared using both the original TRIzol method and the TRIzol method with a commercial clean-up kit. However, the original TRIzol method failed to produce high-quality 2-DE profiles for P. hoffmannianum samples. Among the four modified TRIzol methods, only the TRIzol method with a commercial clean-up kit could yield substantially improved high-quality 2-DE profiles for P. hoffmannianum samples. This combination of the conventional TRIzol method with a commercial clean-up kit potentially represents a promising protein extraction methodology for obtaining high-quality 2-DE profiles for difficult dinoflagellate samples. View Full-Text
Keywords: dinoflagellates; protein preparation; TRIzol reagent; two-dimensional gel electrophoresis; Prorocentrum dinoflagellates; protein preparation; TRIzol reagent; two-dimensional gel electrophoresis; Prorocentrum
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MDPI and ACS Style

Lee, T.C.-H.; Lai, K.K.-Y.; Kwok, C.S.-N.; Xu, S.J.-L.; Lee, F.W.-F. Comparison of Five TRIzol-Based Protein Preparation Methods for 2-DE Production From Challenging Marine Dinoflagellate Samples: A Case Study on Two Benthic Prorocentrum Species. J. Mar. Sci. Eng. 2020, 8, 363. https://doi.org/10.3390/jmse8050363

AMA Style

Lee TC-H, Lai KK-Y, Kwok CS-N, Xu SJ-L, Lee FW-F. Comparison of Five TRIzol-Based Protein Preparation Methods for 2-DE Production From Challenging Marine Dinoflagellate Samples: A Case Study on Two Benthic Prorocentrum Species. Journal of Marine Science and Engineering. 2020; 8(5):363. https://doi.org/10.3390/jmse8050363

Chicago/Turabian Style

Lee, Thomas C.-H.; Lai, Kaze K.-Y.; Kwok, Celia S.-N.; Xu, Steven J.-L.; Lee, Fred W.-F. 2020. "Comparison of Five TRIzol-Based Protein Preparation Methods for 2-DE Production From Challenging Marine Dinoflagellate Samples: A Case Study on Two Benthic Prorocentrum Species" J. Mar. Sci. Eng. 8, no. 5: 363. https://doi.org/10.3390/jmse8050363

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