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Article Dans Une Revue Proteomes Année : 2018

Evaluation of optimized tube-gel methods of sample preparation for large-scale plant proteomics

Résumé

The so-called tube-gel method is a sample preparation protocol allowing for management of SDS for protein solubilization through in-gel protein trapping. Because of its simplicity, we assumed that once miniaturized, this method could become a standard for large scale experiments. We evaluated the performances of two variants of the miniaturized version of the tube-gel method based on different solubilization buffers (Tris-SDS or urea-SDS). To this end, we compared them to two other digestion methods: (i) liquid digestion after protein solubilization in the absence of SDS (liquid method) and (ii) filter-aided sample preparation (FASP). As large-scale experiments may require long term gel storage, we also examined to which extent gel aging affected the results of the proteomics analysis. We showed that both tube-gel and FASP methods extracted membrane proteins better than the liquid method, while the latter allowed the identification and quantification of a greater number of proteins. All methods were equivalent regarding quantitative stability. However, important differences were observed regarding post-translational modifications. In particular, methionine oxidation was higher with the tube-gel method than with the other methods. Based on these results, and considering time, simplicity, and cost aspects, we conclude that the miniaturized tube-gel method is suitable for sample preparation in the context of large-scale experiments.
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hal-01784903 , version 1 (03-05-2018)

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Thierry Balliau, Melisande Blein-Nicolas, Michel Zivy. Evaluation of optimized tube-gel methods of sample preparation for large-scale plant proteomics. Proteomes, 2018, 6 (1(6)), ⟨10.3390/proteomes6010006⟩. ⟨hal-01784903⟩
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