EPR studies of V-ATPase with spin-labeled inhibitors DCC and Archazolid

• Verfasst von: Gölz, Jan Philipp [VerfasserIn]
• Verfasst von: Bockelmann, Svenja [VerfasserIn]
• Verfasst von: Mayer, Kerstin [VerfasserIn]
• Verfasst von: Steinhoff, Heinz-Jürgen [VerfasserIn]
• Verfasst von: Wieczorek, Helmut [VerfasserIn]
• Verfasst von: Huss, Markus [VerfasserIn]
• Verfasst von: Klare, Johann P. [VerfasserIn]
• Verfasst von: Menche, Dirk [VerfasserIn]
• Titel: EPR studies of V-ATPase with spin-labeled inhibitors DCC and Archazolid
• Titelzusatz: interaction dynamics with proton translocating subunit c
• Verf.angabe: Jan Philipp Gölz, Svenja Bockelmann, Kerstin Mayer, Heinz-Jürgen Steinhoff, Helmut Wieczorek, Markus Huss, Johann P. Klare, and Dirk Menche
• Jahr: 2016
• Jahr des Originals: 2015
• Umfang: 9 S.
• Fussnoten: First published: 10 December 2015 ; Gesehen am 02.07.2020
• Titel Quelle: Enthalten in: ChemMedChem
• Ort Quelle: Weinheim [u.a.] : Wiley-VCH, 2006
• Jahr Quelle: 2016
• Band/Heft Quelle: 11(2016), 4, Seite 420-428
• ISSN Quelle: 1860-7187
• DOI: doi:10.1002/cmdc.201500500
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: archazolid
• Sach-SW: EPR spectroscopy
• Sach-SW: natural products
• Sach-SW: spin labels
• Sach-SW: V-ATPase
• K10plus-PPN: 1703397312

Abstract

Abstract Vacuolar-type H+-ATPases (V-ATPases) have gained recent attention as highly promising anticancer drug targets, and therefore detailed structural analyses and studies of inhibitor interactions are very important research objectives. Spin labeling of the V-ATPase holoenzyme from the tobacco hornworm Manduca sexta and V-ATPase in isolated yeast (Saccharomyces cerevisiae) vacuoles was accomplished by two novel methods involving the covalent binding of a (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO) derivative of N,N?-dicyclohexylcarbodiimide (DCC) to the essential glutamate residue in the active site and the noncovalent interaction of a radical analogue of the highly potent inhibitor archazolid, a natural product from myxobacteria. Both complexes were evaluated in detail by electron paramagnetic resonance (EPR) spectroscopic studies and double electron?electron resonance (DEER) measurements, revealing insight into the inhibitor binding mode, dynamics, and stoichiometry as well as into the structure of the central functional subunit?c of these medicinally important hetero-multimeric proton-translocating proteins. This study also demonstrates the usefulness of natural product derived spin labels as tools in medicinal chemistry.