18 Questions
What is the basis for classifying the cell cycle state in the given experiment?
DNA content
Why is Nocodazole block considered efficient in synchronizing cells?
It interferes with microtubule polymerization
What was the threshold used for subsequent analysis in the experiment?
4-fold change in protein abundance
How was protein abundance monitored in Experiment 1?
Using mass spectrometry
Which technique was used to obtain information from the 6 stages of the cell cycle?
SILAC peptide triplet
What was the purpose of quantifying cell cycle markers in different cell populations?
To understand changes in protein abundance throughout the cell cycle
What is the purpose of SILAC labeling in the context of the study mentioned?
To quantify protein expression levels across different stages of the cell cycle.
Which statement accurately explains the difference between the high-turnover and low-turnover proteins in terms of H/L ratios?
The H-concentration of high-turnover protein increases over time.
In the context of the study, what was the role of thymidine in HeLa cell synchronization?
Blocking cells from entering the S phase of the cell cycle.
Why were 3 different isotopic forms of arginine and lysine used for SILAC labeling in HeLa cells?
To allow differentiation between 3 distinct cell populations.
Which technique was used to quantify protein expression levels in the study discussed?
Mass Spectrometry
What was the role of nocodazole in the study involving HeLa cells?
To block entry into a specific phase of the cell cycle.
What is the significance of low occupancy phosphorylation of a protein?
It indicates that only a fraction of the protein copies are phosphorylated at any given time.
What technique was used to analyze all proteins modified by phosphorylation during different stages of the cell cycle in human HeLa cells?
Mass spectrometry
What does SILAC stand for in the context of protein quantification?
Stable Isotope Labeling by Amino Acids in Cell Culture
What happens when non-labelled cells are transferred to heavy SILAC growth medium during protein turnover quantification?
Newly synthesized proteins remain in light form and preexisting proteins incorporate heavy label.
How many phosphorylated proteins were identified during the analysis of the phospho-proteome in human HeLa cells?
6695
What proportion of all proteins was found to be phosphorylated based on the findings presented?
~70%
Learn about SILAC and mass spectroscopy signals with a focus on high-turnover and low-turnover proteins. Understand the principles of H/L ratios of individual proteins, mass spectra of peptides, and how H-concentration affects protein turnover.
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