Dataset Information

The Relationship between Effective Molarity and Affinity Governs Rate Enhancements in Tethered Kinase-Substrate Reactions.

ABSTRACT: Scaffold proteins are thought to accelerate protein phosphorylation reactions by tethering kinases and substrates together, but there is little quantitative data on their functional effects. To assess the contribution of tethering to kinase reactivity, we compared intramolecular and intermolecular kinase reactions in a minimal model system. We found that tethering can enhance reaction rates in a flexible tethered kinase system and that the magnitude of the effect is sensitive to the structure of the tether. The largest effective molarity we obtained was ∼0.08 μM, which is much lower than the effects observed in small molecule model systems and other tethered protein reactions. We further demonstrated that the tethered intramolecular reaction only makes a significant contribution to the observed rates when the scaffolded complex assembles at concentrations below the effective molarity. These findings provide a quantitative framework that can be applied to understand endogenous protein scaffolds and engineer synthetic networks.

SUBMITTER: Speltz EB

PROVIDER: S-EPMC7328773 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Similar Datasets

Project description:Pigments from wood-decay fungi (specifically spalting fungi) have a long history of use in wood art, and have become relevant in modern science due to their longevity and colorfastness. They are presently under investigation as colorants for wood, bamboo, oils, paints and textiles. Major hurdles to their commercialization have been color repeatability (in that the same strain of the same species of fungus may produce different colors over time), and the binding of the pigments to glass storage containers. This is persistent as they do not naturally exist in a loose form. Due to these issues, the 'standard' color for each was historically determined not by the amount of pigment, but by the color in a solution of dichloromethane (DCM), using the CIE L*a*b colorspace. This method of standardization severely limited the use of these pigments in industrial applications, as without a dry form, standard methodologies for repeatable color processing into other materials could not be easily implemented. Recent studies have developed a method to crystalize the red pigment from Scytalidium cuboideum (Sacc. & Ellis) Sigler & Kang, producing a highly pure (99%) solid crystal named 'Dramada'. Herein a method is detailed to compare the molarity of this crystallized pigment to variations in the color, to determine a color saturation curve (by weight) for the pigment from S. cuboideum in DCM and acetone. The molarities for this experiment ranged from 0.024 mM to 19 mM. Each molarity was color read and assigned a CIEL*a*b* value. The results showed that there was a correlation between the molarity and color difference, with the maximum red color occurring between 0.73 mM and 7.3 mM in DCM and between 0.97 mM to 0.73 mM in acetone. Extremely low molarities of pigment produced strong coloration in the solvent, and changes in molarity significantly affected the color of the solution. Having a saturation and color curve for the crystal 'Dramada' from S. cuboideum will allow for the reliable production of distinct colors from a known quantity (by weight) of pigment, erasing the final hurdle towards commercial development of the crystallized pigment from S. cuboideum as an industrial dyestuff.

Dataset Information

The Relationship between Effective Molarity and Affinity Governs Rate Enhancements in Tethered Kinase-Substrate Reactions.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets