Enzymes: from the test tube into the cell


Project leader


Funding source

Swedish Research Council - Vetenskapsrådet (VR)


Project Details

Start date: 01/01/2016
End date: 31/12/2019
Funding: 3200000 SEK


Description

Enzymes are key players in all cellular processes in every living organism. In spite of a wealth of knowledge, the functions of enzymes are still incompletely understood. Numerous application in biotechnology are based on enzymology and approximately half of all pharmaceutical drugs are directed against enzymes, but the design of enzymes for new purposes still presents a challenge and the interactions of targeted enzymes with the components in the intracellular milieu need further studies. Glutathione transferases (GSTs) are ubiquitous detoxication enzymes with multiple cellular functions and are excellently suited to explore various current aspects of enzymology. Genes encoding GSTs will be synthesized, enzymes with enhanced properties will be produced via information-optimized infolog mutants, and the functions of the enzymes characterized. Fluorophore-labeled GSTs as well as fluorogenic substrates will enable single-molecule characterization of the interactions of GSTs with alternative substrates as well as inhibitors, both in vitro and in living cells. Insertion of GST proteins via endocytosis or via transfection from eukaryote expression vectors perturbs components of hormone signaling and studies of proteomes as well as single-cell transcriptomes, and we will explore these newly discovered phenomena in human cells. At the multicellular organism level, model studies will be performed with transgenic GSTs expressed in the fruit fly Drosophila melanogaster and in the thale cress Arabidopsis thaliana. In fruit flies the magnitude of the GST activity in relation to apoptosis and cell death under conditions of chemical stress will be explored, since there are indications that excessive activity may shorten the life-span. In Arabidopsis the utility of GSTs in the bioremediation of soil contaminated with explosives and other toxic agents will be demonstrated. In parallel, poplar GSTs are redesigned via computational predictions and mutagenesis for the ultimate goal of developing fast-growing trees for large-scale applications. In summary, the various functions of enzymes will be illustrated at all levels of complexity ranging from single-molecule kinetics, via enzyme evolution, intracellular effects on molecular networks, and their importance for longevity and biotechnical applications.


Last updated on 2017-28-07 at 08:08