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    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

ATP7B - Kinetics of different ATPases

Cu ATPase Figure 2   different Cu ATPasesIcon N1   SURFE2R N1 data and applications:

We compared the kinetics of the transient currents observed for three different ATPases: The current traces of the Cu2+ ATPases ATP7B and LpCopA show only slow decay times, while the current trace for SERCA decays much faster.

Data from Tadini-Buoninsegni et al, 2017

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Background and Assay Description:
The authors performed 100 μM ATP concentration jumps on samples containing the Ca2+-ATPase SERCA and the Cu2+-ATPases ATP7B and LpCopA. The measurement solutions contained 10 μM free Ca2+ for the SERCA assay and 5 μM CuCl2 for the ATP7B and LpCopA assays.

Impact of the data:
By fitting the current transient decay with a first order exponential decay function, we determined the charge transfer decay time constants (t) for the current signals generated by the various enzymes. It appears that ATP-induced charge transfer in LpCopA (t value of 72 ms) occurs faster than that observed in ATP7B (t value of 140 ms). However, the ATP-induced copper translocation in human as well as bacterial Cu2+-ATPases is much slower than ATP-dependent calcium displacement in SERCA (t value of 25 ms).

Author: Francesco Tadini-Buoninsegni, University of Florence
Target: ATP7B, LpCopA, SERCA
Family / Type: p-type ATPases
Mode of transport: ATP-driven Cu2+ and Ca2+ transport
Organism: Human, Legionella pneumophila
Sample: COS-1 microsomes (ATP7B, SERCA), E. coli membrane fragments (LpCopA)
Expression: Recombinant expression in COS-1 cells (ATP7B, SERCA) or Escherichia coli (LpCopA)
Platform:  SURFE2R One (a predecessor model of the Icon N1   SURFE²R N1)

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