ArticleBiophysical journal2013
Quantitative measurement of protein relocalization in live cells.
Article in Biophysical journal, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 19 citations in OpenAlex.
- Negative feedback-loop mechanisms regulating HOG- and pheromone-MAPK signaling in yeast.Current genetics · 2020Review
- MAPK modulation of yeast pheromone signaling output and the role of phosphorylation sites in the scaffold protein Ste5.Molecular biology of the cell · 2019Article
- CDK and MAPK Synergistically Regulate Signaling Dynamics via a Shared Multi-site Phosphorylation Region on the Scaffold Protein Ste5.Molecular cell · 2018Article
- Spatial and temporal signal processing and decision making by MAPK pathways.The Journal of cell biology · 2017Review
- Yeast GPCR signaling reflects the fraction of occupied receptors, not the number.Molecular systems biology · 2016Article
- Push-Pull and Feedback Mechanisms Can Align Signaling System Outputs with Inputs.Cell systems · 2016Article
- Using measures of single-cell physiology and physiological state to understand organismic aging.Aging cell · 2016Review
- Utilization of extracellular information before ligand-receptor binding reaches equilibrium expands and shifts the input dynamic range.Proceedings of the National Academy of Sciences of the United States of America · 2014Article
- Measuring and sorting cell populations expressing isospectral fluorescent proteins with different fluorescence lifetimes.PloS one · 2014Article
- Automated quantification of the subcellular localization of multicompartment proteins via Q-SCAn.Traffic (Copenhagen, Denmark) · 2013Article
- Feedforward regulation ensures stability and rapid reversibility of a cellular state.Molecular cell · 2013Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
Microscope cytometry provides a powerful means to study signaling in live cells. Here we present a quantitative method to measure protein relocalization over time, which reports the absolute fraction of a tagged protein in each compartment. Using this method, we studied an essential step in the early propagation of the pheromone signal in Saccharomyces cerevisiae: recruitment to the membrane of the scaffold Ste5 by activated Gβγ dimers. We found that the dose response of Ste5 recruitment is graded (EC50 = 0.44 ± 0.08 nM, Hill coefficient = 0.8 ± 0.1). Then, we determined the effective dissociation constant (K(de)) between Ste5 and membrane sites during the first few minutes when the negative feedback from the MAPK Fus3 is first activated. K(de) changed during the first minutes from a high affinity of < 0.65 nM to a steady-state value of 17 ± 9 nM. During the same period, the total number of binding sites decreased slightly, from 1940 ± 150 to 1400 ± 200. This work shows how careful quantification of a protein relocalization dynamic can give insight into the regulation mechanisms of a biological system.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.