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Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Identification of protein complexes with quantitative proteomics in S. cerevisiae

Identification of protein complexes with quantitative proteomics in S. cerevisiae

Published on: March 4, 2009

Protocol for cell image-based spatiotemporal proteomics in budding yeast.

Athanasios Litsios¹, Myra Paz David Masinas¹, Helena Friesen¹

¹Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON M5S 3E1, Canada.

|January 17, 2025

Summary

This study introduces a live-cell imaging protocol to track protein changes throughout the budding yeast cell division cycle. This method enables spatiotemporal proteome analysis for studying cell cycle transitions.

Keywords:

Cell Biology Microscopy Single Cell Systems biology

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Area of Science:

Cell Biology
Biochemistry
Microscopy

Background:

The cell division cycle in eukaryotes involves dynamic changes in protein localization and abundance.
These protein dynamics are crucial for regulating key transitions during cell division.

Purpose of the Study:

To present a detailed protocol for spatiotemporal proteome analysis during the budding yeast cell division cycle.
To enable researchers to visualize and quantify protein behavior in live cells.

Main Methods:

Development of a live-cell imaging protocol.
Strain construction and cultivation of budding yeast.
Microscopy and subsequent image analysis for proteomic data.

Main Results:

A comprehensive protocol for spatiotemporal proteome analysis is described.
The protocol allows for the study of protein dynamics during the cell cycle.
Adaptability of the protocol for various genetic and environmental conditions is highlighted.

Conclusions:

The presented protocol facilitates in-depth analysis of the proteome during the cell division cycle.
This method provides a valuable tool for understanding cell cycle regulation in budding yeast.
The protocol's versatility extends its application to diverse research scenarios.