TY - JOUR
T1 - Characterization of the type 2 NADH:menaquinone oxidoreductases from Staphylococcus aureus and the bactericidal action of phenothiazines
AU - Schurig-Briccio, Lici A.
AU - Yano, Takahiro
AU - Rubin, Harvey
AU - Gennis, Robert B.
N1 - Funding Information:
We thank Dr. S. Monecke (Technical University of Dresden, Germany) for providing the S. aureus RF122 genomic DNA, Dr. H. Nikaido (University of California, Berkeley) for the pRARE2-Km plasmid, Dr. G.N. Bennett (Rice University, Houston) for the GNB10608 strain and Dr. R.K. Jayaswal (Illinois State University) for the Newman strain. We thank all the lab members for their help and useful discussions, and Dr. Ash Pawate for proofreading the text. This work was supported by grants from the National Institutes of Health , GM095600 (RBG), HL16101 (RBG) and AI068942 (HR), and the Global Alliance for TB Drug Development (HR).
PY - 2014/7
Y1 - 2014/7
N2 - Methicillin-resistant Staphylococcus aureus (MRSA) is currently one of the principal multiple drug resistant bacterial pathogens causing serious infections, many of which are life-threatening. Consequently, new therapeutic targets are required to combat such infections. In the current work, we explore the type 2 Nicotinamide adenine dinucleotide reduced form (NADH) dehydrogenases (NDH-2s) as possible drug targets and look at the effects of phenothiazines, known to inhibit NDH-2 from Mycobacterium tuberculosis. NDH-2s are monotopic membrane proteins that catalyze the transfer of electrons from NADH via flavin adenine dinucleotide (FAD) to the quinone pool. They are required for maintaining the NADH/Nicotinamide adenine dinucleotide (NAD+) redox balance and contribute indirectly to the generation of proton motive force. NDH-2s are not present in mammals, but are the only form of respiratory NADH dehydrogenase in several pathogens, including S. aureus. In this work, the two putative ndh genes present in the S. aureus genome were identified, cloned and expressed, and the proteins were purified and characterized. Phenothiazines were shown to inhibit both of the S. aureus NDH-2s with half maximal inhibitory concentration (IC50) values as low as 8 μM. However, evaluating the effects of phenothiazines on whole cells of S. aureus was complicated by the fact that they are also acting as uncouplers of oxidative phosphorylation. This article is part of a Special Issue entitled: 18th European Bioenergetic Conference.
AB - Methicillin-resistant Staphylococcus aureus (MRSA) is currently one of the principal multiple drug resistant bacterial pathogens causing serious infections, many of which are life-threatening. Consequently, new therapeutic targets are required to combat such infections. In the current work, we explore the type 2 Nicotinamide adenine dinucleotide reduced form (NADH) dehydrogenases (NDH-2s) as possible drug targets and look at the effects of phenothiazines, known to inhibit NDH-2 from Mycobacterium tuberculosis. NDH-2s are monotopic membrane proteins that catalyze the transfer of electrons from NADH via flavin adenine dinucleotide (FAD) to the quinone pool. They are required for maintaining the NADH/Nicotinamide adenine dinucleotide (NAD+) redox balance and contribute indirectly to the generation of proton motive force. NDH-2s are not present in mammals, but are the only form of respiratory NADH dehydrogenase in several pathogens, including S. aureus. In this work, the two putative ndh genes present in the S. aureus genome were identified, cloned and expressed, and the proteins were purified and characterized. Phenothiazines were shown to inhibit both of the S. aureus NDH-2s with half maximal inhibitory concentration (IC50) values as low as 8 μM. However, evaluating the effects of phenothiazines on whole cells of S. aureus was complicated by the fact that they are also acting as uncouplers of oxidative phosphorylation. This article is part of a Special Issue entitled: 18th European Bioenergetic Conference.
KW - Bioenergetics/electron transfer complex
KW - Enzyme inhibitor
KW - NADH dehydrogenase
KW - Phenothiazine
KW - Respiratory chain
KW - Staphylococcus aureus
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U2 - 10.1016/j.bbabio.2014.03.017
DO - 10.1016/j.bbabio.2014.03.017
M3 - Article
C2 - 24709059
AN - SCOPUS:84901820211
SN - 0005-2728
VL - 1837
SP - 954
EP - 963
JO - Biochimica et Biophysica Acta - Bioenergetics
JF - Biochimica et Biophysica Acta - Bioenergetics
IS - 7
ER -