Combinatorial Chemical Reengineering of the Alpha Class Glutathione Transferases
2004 (English)In: Bioconjugate Chemistry, ISSN 1043-1802 (print) 1520-4812 (online), Vol. 15, no 4, 718-727 p.Article in journal (Refereed) Published
Previously, we discovered that human glutathione transferases (hGSTs) from the alpha class can be rapidly and quantitatively modified on a single tyrosine residue (Y9) using thioesters of glutathione (GS-thioesters) as acylating reagents. The current work was aimed at exploring the potential of this site-directed acylation using a combinatorial approach, and for this purpose a panel of 17 GS-thioesters were synthesized in parallel and used in screening experiments with the isoforms hGSTs A1-1, A2-2, A3-3, and A4-4. Through analytical HPLC and MALDI-MS experiments, we found that between 70 and 80% of the reagents are accepted and this is thus a very versatile reaction. The range of ligands that can be used to covalently reprogram these proteins is now expanded to include functionalities such as fluorescent groups, a photochemical probe, and an aldehyde as a handle for further chemical derivatization. This site-specific modification reaction thus allows us to create novel functional proteins with a great variety of artificial chemical groups in order to, for example, specifically tag GSTs in biological samples or create novel enzymatic function using appropriate GS-thioesters.
Place, publisher, year, edition, pages
2004. Vol. 15, no 4, 718-727 p.
IdentifiersURN: urn:nbn:se:liu:diva-13196DOI: 10.1021/bc034192+OAI: oai:DiVA.org:liu-13196DiVA: diva2:18020