Nanion Technologies
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Data and Applications


Action of NaK ATPase

Surfer_NaK_ATPase

The Sodium-Potassium ATPase, also known as Na+/K+ pump is responsible for the active transport of Na+ and K+ in the cells containing relatively high concentrations of K+ ions but low concentrations of Na+ ions. The Na+/K+-ATPase helps maintain resting potential, avail transport, and regulate cellular volume. Here, we demonstrate the conductance of the pump, in the presence of ATP, Na+ and K+, or in the presence of ATP and Na+.

Adenine nucleotide translocator recorded on the Surfer

Surfer_ANT_trace

The Adenine Nucleotide Translocator (ANT) is located in the inner membrane of mitochondria and transports, as the name suggests, either ADP or ATP alone or against each other.  Recordings were done on mitochondria inner membrane preparations from pigs heart.  Example traces display the action of ANT either in the presence of ATP, in an other experiment in the presence of ADP.

PepT1 on SURFE2R N96

Surfer N96 PepT1 on 96 well plate

Graphical user interface of screening software used on the SURFE2R N96. Traces of PepT1 recorded from membrane preparations of PepT1 overexpressing HEK293 cells are presented. The PepT1 signal is evoked by 20mM glycylglycin.

Pharmacology on PepT1

Surfer N96 PepT1 Losarthan IC50 datacontrol

Transport by PepT1 is inhibited by the kompetitive antagonist Losartan. When activated with 20mM Gly-Gly, the IC50 of Losartan was 80.4 µM. Data were analysed with DataControl - the Analysis software tool for recordings on the SURFE2R N96 and SyncroPatch96/384PE.

Respiratory Chain Complex I and III

Surfer_RC_complexI_III_trace

In the respiratory chain Complex I and III, oxidation of NADH leads to transport of 4 protons across the membrane. The electrons are transferred to cytochrome c. Because of the electron leakage to oxygen, both complex are the main sites of production of harmful superoxide. The Surfer-experiments, application of 100µM NADH results in a current over the mitochondrial membrane, where Complex I and III are expressed.

Respiratory Chain Complex II and III

Surfer_RC_II_III

Respiratory Chain Complex II is a succinate dehydrogenase. In this complex, succinate is oxidated to fumarate. The electrons are transfered to the quinone pool, the Complex III (cytochrome bc1 complex) and cytochrome c, while four protons are transported to the intermembrane space.

Respiratory Chain Complex IV

Surfer_RC_IV

The Respiratory Chain Complex IV, known as Cytochrom c oxidase, reduces oxygen by cunsumption of electrons from cytocrome c and transport of two protons to the intermembrans space. Here the reaction runs backwards.

Respiratory Chain Complex V ATPSyntase

Surfer_RC_V

The Respiratory Chain Complex V uses the transmembrane proton gradient (produced by Complex I, III and IV) to generates ATP from ADP plus phosphate. One component of ATP synthase acts as an ion channel that provides for a proton flux back into the mitochondrial matrix. This reflux releases free energy, which is used to drive ATP synthesis, catalyzed by the other component of the complex. Experiments on the Surfer show the action of the ATPase after application of ATP.

Surfer Recordings of PepT1

Surfer_PepT1_traces_IC50

Representive traces of PepT1 overexpressed in CHO cells. PepT1 (SLC15A1) is expressed mainly in intestine and kidney, it enables uptake of oligopeptides. The PepT1 signal is evoked by glycylglycin. The transport of oligopeptides can be inhibited by addition of Lys[Z(NO2)]-Val. The IC50 of Lys[Z(NO2)]-Val was 380µM.