S-Nitrosation of monocarboxylate transporter 1: inhibition of pyruvate-fueled respiration and proliferation of breast cancer cells.
Energy substrates metabolized through mitochondria (e.g., pyruvate, glutamine) are required for biosynthesis of macromolecules in proliferating cells. Because several mitochondrial proteins are known to be targets of S-nitrosation, we determined whether bioenergetics are modulated by S-nitrosation and defined the subsequent effects on proliferation. The nitrosating agent S-nitroso-L-cysteine (L-CysNO) was used to initiate intracellular S-nitrosation, and treatment decreased mitochondrial function and inhibited proliferation of MCF7 mammary adenocarcinoma cells. Surprisingly, the d-isomer of CysNO (D-CysNO), which is not transported into cells, also caused mitochondrial dysfunction and limited proliferation. Both L- and D-CysNO also inhibited cellular pyruvate uptake and caused S-nitrosation of thiol groups on monocarboxylate transporter 1, a proton-linked pyruvate transporter. These data demonstrate the importance of mitochondrial metabolism in proliferative responses in breast cancer and highlight a novel role for inhibition of metabolic substrate uptake through S-nitrosation of exofacial protein thiols in cellular responses to nitrosative stress.
Department of Biophysics, Medical College of Wisconsin, Milwaukee, WI 53226, USA; Redox Biology Program, Medical College of Wisconsin, Milwaukee, WI 53226, USA. Electronic address: annerdiers@gmail.com.
Anne R
AR
Diers
Department of Biophysics, Medical College of Wisconsin, Milwaukee, WI 53226, USA; Redox Biology Program, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
Katarzyna A
KA
Broniowska
Department of Biophysics, Medical College of Wisconsin, Milwaukee, WI 53226, USA; Redox Biology Program, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
Ching-Fang
CF
Chang
Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
R Blake
RB
Hill
Department of Biophysics, Medical College of Wisconsin, Milwaukee, WI 53226, USA; Redox Biology Program, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
Neil
N
Hogg
D049109:Cell Proliferation / Q000187:drug effects; D019069:Cell Respiration / Q000187:drug effects / Q000235:genetics; D003545:Cysteine / Q000008:administration & dosage / Q000031:analogs & derivatives; D006801:Humans; D061986:MCF-7 Cells; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D027501:Monocarboxylic Acid Transporters / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D015538:Nitrosation; D019289:Pyruvic Acid / Q000378:metabolism*; D026403:S-Nitrosothiols / Q000008:administration & dosage; D013438:Sulfhydryl Compounds / Q000378:metabolism; D027981:Symporters / Q000378:metabolism*; D000099233:Monocarboxylate Transport Protein 1
D003545:Cysteine; D027501:Monocarboxylic Acid Transporters; D009569:Nitric Oxide; D019289:Pyruvic Acid; D026403:S-Nitrosothiols; D013438:Sulfhydryl Compounds; D027981:Symporters; D000099233:Monocarboxylate Transport Protein 1; C021315:S-nitrosocysteine
Extracellular flux analysis; Free radicals; Nitric oxide; Nitrosylation; Redox; Thiol