Evaluation of protein preparations from bacteria using proteomics approaches: Application to an Antarctic bacterium Pseudomonas syringae
Ehab F. Abou-Eladab 1, 2*, M Mohammed Idris2, Medicharla V Jagannadham2
1Mansoura University, Faculty of Specific Education-Damietta, 34517 New Damietta City, Damietta, Egypt
2 Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500007, India.
Abstract: The proteins of Pseudomonas syringae pv. TomatoT1 were prepared using a protein preparation method with some modifications and applied the procedure to an Antarctic Pseudomonas syringae Lz4W and evaluate the results using proteomics methods. The preparation of proteins was studied using Pseudomonas syringae pv. tomato T1 as a model bacterium and separated the proteins on SDS-PAGE. Suitable method is applied for the preparation of proteins from Pseudomonas syringae Lz4W. The gel bands cut into small slices, in-gel digested with trypsin and the resulting peptide mixtures analyzed by proteomics analysis. Our results showed a total of 190 proteins were identified from supernatant and 326 proteins from pellet when bacteria grown at 22oC by using Method A. Also Pseudomonas syringae pv. tomato T1 showed a total of 301 proteins grown at 28oC by using Method B, whereas only 33 proteins were identified using MALDI. The proteomic approaches used in the present study are very useful to evaluate the efficiency of the protein preparation methods. This study highlights that the protein preparation method plays a critical role in proteomics studies of bacteria.
[Ehab F. Abou-Eladab, M Mohammed Idris, Medicharla V Jagannadham. Evaluation of protein preparations from bacteria using proteomics approaches: Application to an Antarctic bacterium Pseudomonas syringaeNature and Science 2012; 10(4): 102-114]. (ISSN: 1545-0740). http://www.sciencepub.net/nature. 16
Keywords: Proteins of Pseudomonas syringae
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