Evaluation of four different strategies to characterize plasma membrane proteins from banana roots

Ciência e Agrotecnologia

Suzana Antunes Lourençoni Garcia²  Bart Panis¹  Rony Swennen²  Sebastien Christian Carpentier² 

[1] ,KU Leuven Division of Crop Biotechnics Leuven,Belgium

关键词: Aqueous-polymer two-phase system technique;    microsomal and full proteome fractions;    protein identification;    Sistema aquoso de duas fases;    frações microssomal e de proteoma total;    identificação de proteínas;   

DOI  :  10.1590/S1413-70542014000500002

来源: SciELO

【 摘 要 】

Plasma membrane proteins constitute a very important class of proteins. They are involved in the transmission of external signals to the interior of the cell and selective transport of water, nutrients and ions across the plasma membrane. However, the study of plasma membrane proteins is challenging because of their poor solubility in aqueous media and low relative abundance. In this work, we evaluated four different strategies for the characterization of plasma membrane proteins from banana roots: (i) the aqueous-polymer two-phase system technique (ATPS) coupled to gelelectrophoresis (gel-based), and (ii) ATPS coupled to LC-MS/MS (gel free), (iii) a microsomal fraction and (iv) a full proteome, both coupled to LC-MS/MS. Our results show that the gel-based strategy is useful for protein visualization but has major limitations in terms of time reproducibility and efficiency. From the gel-free strategies, the microsomal-based strategy allowed the highest number of plasma membrane proteins to be identified, followed by the full proteome strategy and by the ATPS based strategy. The high yield of plasma membrane proteins provided by the microsomal fraction can be explained by the enrichment of membrane proteins in this fraction and the high throughput of the gel-free approach combined with the usage of a fast high-resolution mass spectrometer for the identification of proteins.

【 授权许可】

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