Protein Proteomics – Abstract

Proteomics is an important field of study, because the analysis of protein interactions and protein structure is critical to understanding protein function.  Modifications in the protein, especially post-translational modifications, are key to understanding function and roles of the proteins, which is why there is a need in biochemistry to sequence proteins.  In this research experiment, E. Coli bacterial cells are being lytically infected with the t7 phage.The lytic cycle starts when the bacteriophage infects the cell, and the phage DNA then replicates and creates phage protein in the E. Coli cell.2   My Honors project will be conducted in close collaboration with Professor Williamson’s lab.  E. coli bacterial cells will be infected with the t7 phage at various times (5, 25, 45 and 65 minutes), and we will be looking for the differences in the proteins expressed based on the different time exposure of the t7 phage.  By looking for modifications in the proteins on the different time exposure, protein structure and function can then be deduced.  The procedure includes growing the cell cultures, extracting and separating the proteins, and then analyzing the peptide fragments using HPLC-ESI-MS/MS and the protein database SEQUEST.  The importance of this project is that we are using a proteomics procedure that uses a HPLC-ESI-MS/MS (high pressure liquid chromatography-electrospray ionization-tandem mass spectrometer) method.  Mass spectrometry has emerged as a core tool for protein analysis and sequencing, especially since most mass spectrometers have advanced in resolution, mass accuracy and sensitivity.3,4  As a time and money efficient tool, mass spectrometry is important in the field of proteomics, and with this Honors project we are trying practice bioanalysis using a HPLC-ESI-MS/MS method.

 

Works Cited

  1. Bartel, Paul L., Jennifer A. Roecklein, Dhruba Sengupta, and Stanley Fields. “A protein linkage map of Escherichia coli bacteriophage T7.” Nature Genetics 12, no. 1 (1996): 72-77. Accessed February 26, 2017. doi:10.1038/ng0196-72.
  2. “Lytic Cycle.” Lytic Cycle. Accessed February 26, 2017. http://sites.fas.harvard.edu/~biotext/animations/lyticcycle.html.
  3. Gundry, Rebekah L., Melanie Y. White, Christopher I. Murray, Lesley A. Kane, Qin Fu, Brian A. Stanley, and Jennifer E. Van Eyk. “Preparation of Proteins and Peptides for Mass Spectrometry Analysis in a Bottom-Up Proteomics Workflow.” Current Protocol of Molecular Biology, July 19, 2010. Accessed February 26, 2017. doi: 10.1002/0471142727.mb1025s88.
  4. Aebersold, Reudi, and Matthias Mann. “Mass spectrometry-based proteomics.” Nature 422 (March 13, 2003). Accessed February 26, 2017. doi:10.1038/nature01511.

Comments

  1. Anna Mika says:

    Sounds interesting. Keep us posted about the progress of the project.

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