Reference: Gencoglu M, et al. (2017) Measurement of In Vivo Protein Binding Affinities in a Signaling Network with Mass Spectrometry. ACS Synth Biol 6(7):1305-1314

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Abstract


Protein interaction networks play a key role in signal processing. Despite the progress in identifying the interactions, the quantification of their strengths lags behind. Here we present an approach to quantify the in vivo binding of proteins to their binding partners in signaling-transcriptional networks, by the pairwise genetic isolation of each interaction and by varying the concentration of the interacting components over time. The absolute quantification of the protein concentrations was performed with targeted mass spectrometry. The strengths of the interactions, as defined by the apparent dissociation constants, ranged from subnanomolar to micromolar values in the yeast galactose signaling network. The weak homodimerization of the Gal4 activator amplifies the signal elicited by glucose. Furthermore, combining the binding constants in a feedback loop correctly predicted cellular memory, a characteristic network behavior. Thus, this genetic-proteomic binding assay can be used to faithfully quantify how strongly proteins interact with proteins, DNA and metabolites.

Reference Type
Journal Article
Authors
Gencoglu M, Schmidt A, Becskei A
Primary Lit For
GAL3 | GAL1 | GAL4 | GAL80 | GAL3-GAL80 transcription regulation complex | GAL1-GAL80 transcription regulation complex | GAL4-GAL80 transcription repressor complex
Additional Lit For
GAL7

Interaction Annotations


Genetic Interactions

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Interactor Interactor Allele Assay Annotation Action Phenotype SGA score P-value Source Reference

Physical Interactions 5 entries for 4 genes

Increase the total number of rows showing on this page by using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i" buttons located within a cell for an annotation to view further details about experiment type and any other genes involved in the interaction.

InteractorInteractorAssayAnnotationActionModification

PCA is the best fit system for the genetic-proteomic binding assay introduced in this paper

GAL1GAL80PCAmanually curatedBait-HitNo Modification

PCA is the best fit system for the genetic-proteomic binding assay introduced in this paper

GAL3GAL80PCAmanually curatedBait-HitNo Modification

PCA is the best fit system for the genetic-proteomic binding assay introduced in this paper

GAL4GAL4PCAmanually curatedHit-BaitNo Modification

PCA is the best fit system for the genetic-proteomic binding assay introduced in this paper

GAL80GAL4PCAmanually curatedBait-HitNo Modification

PCA is the best fit system for the genetic-proteomic binding assay introduced in this paper

GAL80GAL80PCAmanually curatedHit-BaitNo Modification
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