Related Experiment Video
Updated: Aug 19, 2026

Measuring In Vitro ATPase Activity for Enzymatic Characterization
Published on: August 23, 2016
The thermodynamic efficiency of the Ca2+-Mg2+-ATPase is one hundred percent
Abstract:
The thermodynamic efficiency of the Ca2+ -Mg2+ -ATPase of skeletal sarcoplasmic reticulum has been evaluated by comparing the Ca2+ gradient established with the ATP/(ADP*Pi) ratio. The evaluation was made at an external Ca2+ level (4.7 X 10(-8) M) which is below the Km value of 7 X 10(-8) M. The Mg-ATP and phosphate concentrations were held constant (0.1 mM) and the ADP concentration was varied. Maximal uptake to an internal free Ca2+ concentration of 17 mM was observed at infinite ATP/(ADP*Pi) ratio (absence of ADP). This corresponds to a [Ca2+] i/[Ca2+] 0 gradient of 3.6 X 10(5). A Ca2+ gradient one-half as large was observed at an ATP/(ADP*Pi) ratio of 3.5 X 10(3) M-1. The square of the Ca2+ gradient is shown to be proportional to the ATP/(ADP*Pi) ratio, for finite values of the latter. The proportionality constant is identical to the equilibrium constant for hydrolysis of ATP (9.02 X 10(6) M) under these conditions (0.1 mM Mg2+, 30 degrees C). The intrinsic thermodynamic efficiency of the pump is shown to be 100%, with a maximal uncertainty of 3%. The efficiency is lower under less optimal conditions, when the pump is inhibited and passive leak processes compete.
Related Concept Videos
ATP Synthase: Mechanism
ATP Synthase: Structure
ATP Energy Storage and Release
One example of energy coupling using ATP involves a...
ATP Driven Pumps II: P-type Pumps
A typical P-type pump has three cytosolic domains: nucleotide-binding (N), phosphorylation (P), and activator (A) domains. These domains are connected to the membrane-spanning helices by short amino acid segments. ATP hydrolysis and covalent phosphoenzyme intermediate formation are crucial parts of the catalytic cycle. At the highly...
ATP Driven Pumps III: V-type Pumps
The peripheral or cytosolic V1 domain with eight subunits is involved in ATP hydrolysis. The integral or transmembrane V0 domain containing at least five subunits...
ATP Energy Storage and Release
One example of energy coupling using ATP involves a...

