scholarly journals Regulatory Mechanisms of the Calcium Transport System of Fragmented Rabbit Sarcoplasmic Reticulum

1971 ◽  
Vol 57 (1) ◽  
pp. 64-70 ◽  
Author(s):  
A. Weber
Keyword(s):  
Steady State ◽  
Sarcoplasmic Reticulum ◽  
Transport System ◽  
Rate Constant ◽  
Calcium Transport ◽  
Regulatory Mechanisms ◽  
Low Permeability ◽  
Free Medium ◽  
Hydrolysis Of ◽  
Permeability State

The outflux of calcium from vesicles of sarcoplasmic reticulum is controlled by the concentration of ionized calcium in the medium. In a calcium-free medium calcium outflux is at a minimum provided ATP and Mg are present. If they are removed calcium outflux becomes rapid and has a rate constant similar to that of outflux during steady-state 40Ca-45Ca exchange with saturating calcium concentrations in the medium. In order to maintain the low permeability state of the membrane only the presence but not the continued hydrolysis of ATP is necessary.

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

1987 ◽  
Vol 42 (5) ◽  
pp. 641-652 ◽  
Author(s):  
Wilhelm Hasselbach ◽  
Lore Stephan
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Calcium Transport ◽  
Activation Volume ◽  
Reaction Scheme ◽  
Pressure Effects ◽  
Calcium Dependent ◽  
Nitrophenyl Phosphate ◽  
Hydrolysis Of ◽  
Sarcoplasmic Reticulum Calcium ◽  
Effect Of Hydrostatic Pressure

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.

Presteady-state kinetics of Bacillus 1,3–1,4-β-glucanase: binding and hydrolysis of a 4-methylumbelliferyl trisaccharide substrate

2001 ◽  
Vol 357 (1) ◽  
pp. 195-202
Author(s):  
Mireia ABEL ◽  
Antoni PLANAS ◽  
Ulla CHRISTENSEN
Keyword(s):  
Steady State ◽  
Rate Constant ◽  
Product Formation ◽  
Lag Phase ◽  
Wild Type ◽  
Induced Fit ◽  
Enzyme Substrate ◽  
Kinetics Of ◽  
Hydrolysis Of ◽  
Presteady State Kinetics

In the present study the first stopped-flow experiments performed on Bacillus 1,3–1,4-β-glucanases are reported. The presteady-state kinetics of the binding of 4-methylumbelliferyl 3-O-β-cellobiosyl-β-d-glucoside to the inactive mutant E134A, and the wild-type-catalysed hydrolysis of the same substrate, were studied by measuring changes in the fluorescence of bound substrate or 4-methylumbelliferone produced. The presteady-state traces all showed an initial lag phase followed by a fast monoexponential phase leading to equilibration (for binding to E134A) or to steady state product formation (for the wild-type reaction). The lag phase, with a rate constant of the order of 100s−1, was independent of the substrate concentration; apparently an induced-fit mechanism governs the formation of enzyme–substrate complexes. The concentration dependencies of the observed rate constant of the second presteady-state phase were analysed according to a number of reaction models. For the reaction of the wild-type enzyme, it is shown that the fast product formation observed before steady state is not due to a rate-determining deglycosylation step. A model that can explain the observed results involves, in addition to the induced fit, a conformational change of the productive ES complex into a form that binds a second substrate molecule in a non-productive mode.

The Inhibition of the Calcium Transport ATPase of the Sarcoplasmic Reticulum by Fluorescamine: Evidence for an Oligomeric Functional Unit of the Calcium Transport System

1980 ◽  
Vol 35 (11-12) ◽  
pp. 1005-1011 ◽  
Author(s):  
Wilhelm Hasselbach ◽  
Andrea Migala
Keyword(s):  
Transport System ◽  
Calcium Transport ◽  
Functional Unit ◽  
Amino Group ◽  
Transport Atpase ◽  
Calcium Dependent ◽  
Inhibiting Effect ◽  
The Relationship ◽  
Hydrolysis Of ◽  
Protein Moiety

The labelling of the protein moiety of the sarcoplasmic calcium transport ATPase by fluo- rescamine suppresses calcium transport, calcium dependent ATPase activity, protein phosphoryla­tion by [γ-32P]ATP and [32P]phosphate at different extent of amino group substitution. For the hydrolysis of para nitrophenylphosphate by the calcium transport ATPase it is shown that the relationship between the extent of amino group labelling can considerably be altered by the temperature and the presence of ethyleneglycol. It is shown that the amino residues of the phosphatidylethanolamine moiety do not contribute to the inhibiting effect of fluorescamine labelling. The observations suggest that the different functions of the calcium transport system are based on the cooperation of a varying number of calcium transport ATPase molecules.

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