• SURFE²R N1

    Easy-to-learn all-in-one device, ideal for teaching and university research
  • SURFE²R N1

    Finally label-free functional assays for transporters available
  • SURFE²R N1

    High signal amplification compared to patch-clamp: transport & binding assays
  • SURFE²R N1

    The only instrument on the market for SSM-based electrophysiology
  • SURFE²R N1

    Turn-key system for efficient transporter protein analysis

2009 - Inhibitory effect of Pb2+ on the transport cycle of the Na+,K+-ATPase

Icon N1   SURFE²R ONE (a predecessor model of SURFE²R N1) publication in Chemical Research in Toxicology (2009)

Authors:
Gramigni E., Tadini-Buoninsegni F., Bartolommei G., Santini G., Chelazzi G., Moncelli M.R.

Journal:
Chemical Research in Toxicology (2009) 22(10):1699-1704


Abstract:

The effect of Pb2+ on the transport cycle of the Na+,K+-ATPase was characterized in detail at a molecular level by combining electrical and biochemical measurements. Electrical measurements were performed by adsorbing purified membrane fragments containing Na+,K+-ATPase on a solid-supported membrane. Upon adsorption, the Na+,K+-ATPase was activated by carrying out concentration jumps of different activating substrates, for example, Na+ and ATP. Charge movements following Na+,K+-ATPase activation were measured in the presence of various Pb2+ concentrations to investigate the effect of Pb2+ on different ion translocating steps of the pump cycle. These charge measurements were then compared to biochemical measurements of ATPase activity in the presence of increasing Pb2+ concentration. Our results indicate that Pb2+ inhibits cycling of the enzyme, but it does not affect cytoplasmic Na+ binding and release of Na+ ions at the extracellular side at concentrations below 10 μM. To explain the inhibitory effect of Pb2+ on the Na+,K+-ATPase, we propose that Pb2+ may interfere with the hydrolytic cleavage of the phosphorylated intermediate E2P, which occurs in the K+-related branch of the pump cycle.


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