Direct evidence for the interaction of stigmatellin with a protonated acidic group in the bc(1) complex from Saccharomyces cerevisiae as monitored by FTIR difference spectroscopy and 13C specific labeling

Biochemistry
Michaela RitterPetra Hellwig

Abstract

In this study a combined electrochemical and FTIR spectroscopic approach was applied to monitor the binding of stigmatellin, a Q(o) site inhibitor of the cytochrome bc(1) complex from Saccharomyces cerevisiae. Natural stigmatellin A induced clear shifts in the redox-induced FTIR difference spectra. For data interpretation a stigmatellin derivative (UST) with the conjugated trienes replaced by an aliphatic tail was synthesized, and the carbonyl group shown in crystal structures to interact with His181, the [2Fe-2S] ligand of the Rieske, was specifically (13)C labeled. Electrochemically induced FTIR difference spectra of the inhibitors in CH(3)OD were obtained and revealed signals characteristic for the oxidized and reduced forms of the labeled and unlabeled compounds. On the basis of signals from the inhibitors alone, the binding of the inhibitor to the bc(1) complex was monitored. Direct evidence for the interaction of the carbonyl group with the protein was provided by the observed shift of the nu(C=O) vibrational mode of about 10 cm(-1). In addition, redox-dependent reorganizations of the protein were identified, including protonation changes of acidic residues at 1746 and 1734 cm(-1). The conformational changes observed upon...Continue Reading

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Citations

Aug 15, 2008·The Journal of Biological Chemistry·Buddha GurungChang-An Yu
Apr 29, 2006·Journal of Molecular Microbiology and Biotechnology·Dirk FlemmingThorsten Friedrich
Feb 3, 2005·Yeast
Jul 16, 2014·Biochimica Et Biophysica Acta·Petra Hellwig
Jul 1, 2013·Molecular Informatics·Marta P CarrascoDaniel J V A Dos Santos
Oct 1, 2011·Bioorganic & Medicinal Chemistry·Tiago RodriguesRita C Guedes
Mar 14, 2007·Biochimica Et Biophysica Acta·Astrid R KlingenG Matthias Ullmann
Jul 7, 2005·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·Steven J CohenNeal J Meropol

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