This assay is designed to obtain the in vitro kinetic data of T. brucei recombinant trypanothione synthetase. The enzyme catalyzes the ATP-dependent ligation of spermidine (Spd) and GSH to generate glutathionylspermidine (Gsp) and also of Gsp and GSH to finally produce trypanothione (T(SH)2). The data was obtained in an spectrophotometric assay that links ADP production with NADH consumption through the piruvte kinase and lactate dehydrogenase.
SEEK ID: https://fairdomhub.org/assays/194
Assay type: Experimental Assay Type
Technology type: Enzymatic Activity Measurements
Organisms: No organisms
Created: 27th Nov 2012 at 15:54
Last updated: 8th Nov 2017 at 15:21
The SilicoTryp project aims at the creation of a “Silicon Trypanosome”, a comprehensive, experiment-based, multi-scale mathematical model of trypanosome physiology.
Trypanosomes are blood-stream parasites transmitted by tsetse flies; they cause African sleeping sickness in humans and livestock. Currently available drugs have severe side effects, and the parasites are rapidly developing resistance.
In this project, we collect a wide range of new experimental data on the parasite in its various
Aim. To provide critical quantitative parameter information and to model redox balance by determining the cellular concentration of all enzymes involved in the trypanothione-dependent hydroperoxide detoxification system of trypanosomes and by performing the kinetic characterization of the involved enzymes under pseudo-physiological conditions.
Snapshots: No snapshots
Studies: Determination of the redox state and the total concentration of the tryp..., Kinetic characterization of trypanothione-dependent enzymes, Kinetic modelling of Trypanothione Synthetase to elucidate the enzyme me...
The enzymes involved in the trypanothione metabolism will be studied in a uniform assay medium that mimics the intracellular milieu of the parasite.
Person responsible: Alejandro Leroux
Snapshots: No snapshots
The file contains the initial rate measurements of TbTryS obtained under different substrate and product initial concentrations.
This is a protocol for determining the activity of the T. brucei Trypanothione synthetase under in vivo-like conditions.
This method describes the preparation of the in vivo-like buffer for the measurement of bloodstream T. brucei recombinant enzymes under pseudo-physiological conditions.
Date Published: 3rd Jul 2013
Journal: J. Biol. Chem.
PubMed ID: 23814051