Pressure Effects on the Interactions of the Sarcoplasmic Reticulum Calcium Transport Enzyme with Calcium and para-Nitrophenyl Phosphate

Abstract

The effect of hydrostatic pressure on calcium dependent p-nitrophenyl phosphate hydrolysis of the sarcoplasmic reticulum calcium transport enzyme has been investigated at different degree of enzyme saturation by calcium and Mg-p-nitrophenyl phosphate to distinguish between activation and binding volumes. The enzyme saturated by both ligands displays a significant dependence of the activation volume on pressure, rising from 20 ml/mol at atmospheric pressure (0.1 MPa) to 80 ml/mol at 100 MPa. At subsaturating concentration of Mg-p-nitrophenyl phosphate an activation volume of 35 ml/mol prevails between 0.1 and 40 MPa. At subsaturating concentration of calcium the activation volume approximates 80 ml/mol in the same pressure range. The binding volume for both substrates is likewise pressure dependent falling from 20 ml/mol to 0 ml/mol for Mg-p-nitrophenyl phosphate and rising from 67 ml/mol to 155 ml/mol for calcium. The pressure dependence of activation and binding volumes is analysed on account of a simplified reaction scheme yielding activation volumes and rate constants for individual reaction steps.

Characterizing the dual-wavelength dye indo-1 for calcium-ion sensing under pressure

“…We have neglected, however, to account for a pressuredependent pCa; the dissociation constant of the chelator EGTA (pK EGTA ) depends on pH and pressure. 20,21 We estimate the pH change upon pressurization using…”