Exploring the Arabidopsis Proteome: Influence of Protein Solubilization Buffers on Proteome Coverage

International Journal of Molecular Sciences

Claudius Marondedze¹  Aloysius Wong¹  Arnoud Groen²  Natalia Serrano¹  Boris Jankovic¹  Kathryn Lilley²  Christoph Gehring¹  Ludivine Thomas¹ 

[1] Biological and Environmental Sciences and Engineering Division, 4700 King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi Arabia; E-Mails:;Cambridge Centre for Proteomics, Cambridge System Biology, University of Cambridge, Department of Biochemistry, Tennis Court Road, Cambridge CB2 1QR, UK; E-Mails:

关键词: two-dimensional gel electrophoresis (2DE);    Arabidopsis thaliana;    detergent;    mass spectrometry (MS);    OFFGEL fractionation;   

DOI  :  10.3390/ijms16010857

来源: mdpi

【 摘 要 】

The study of proteomes provides new insights into stimulus-specific responses of protein synthesis and turnover, and the role of post-translational modifications at the systems level. Due to the diverse chemical nature of proteins and shortcomings in the analytical techniques used in their study, only a partial display of the proteome is achieved in any study, and this holds particularly true for plant proteomes. Here we show that different solubilization and separation methods have profound effects on the resulting proteome. In particular, we observed that the type of detergents employed in the solubilization buffer preferentially enriches proteins in different functional categories. These include proteins with a role in signaling, transport, response to temperature stimuli and metabolism. This data may offer a functional bias on comparative analysis studies. In order to obtain a broader coverage, we propose a two-step solubilization protocol with first a detergent-free buffer and then a second step utilizing a combination of two detergents to solubilize proteins.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.