Protein Set for Normalization of Quantitative Mass Spectrometry Data

Autor: Lee, Wooram
Rok vydání: 2014
Předmět:
Druh dokumentu: Diplomová práce
Popis: Mass spectrometry has been recognized as a prominent analytical technique for peptide and protein identification and quantitation. With the advent of soft ionization methods, such as electrospray ionization and matrix assisted laser desorption/ionization, mass spectrometry has opened a new era for protein and proteome analysis. Due to its high-throughput and high-resolution character, along with the development of powerful data analysis software tools, mass spectrometry has become the most popular method for quantitative proteomics. Stable isotope labeling and label-free quantitation methods are widely used in quantitative mass spectrometry experiments. Proteins with stable expression level and key roles in basic cellular functions such as actin, tubulin and glyceraldehyde-3-phosphate dehydrogenase, are frequently utilized as internal controls in biological experiments. However, recent studies have shown that the expression level of such commonly used housekeeping proteins is dependent on cell type, cell cycle or disease status, and that it can change as a result of a biochemical stimulation. Such phenomena can, therefore, substantially compromise the use of these proteins for data validation. In this work, we propose a novel set of proteins for quantitative mass spectrometry that can be used either for data normalization or validation purposes. The protein set was generated from cell cycle experiments performed with MCF-7, an estrogen receptor positive breast cancer cell line, and MCF-10A, a non-tumorigenic immortalized breast cell line. The protein set was selected from a list of 3700 proteins identified in the different cellular sub-fractions and cell cycle stages of MCF-7/MCF-10A cells, based on the stability of spectral count data (CV
Master of Science
Databáze: Networked Digital Library of Theses & Dissertations
Externí odkaz: