| Effect of pressure and calcium on the reversible inhibition of the sarcoplasmic-reticulum calcium-transport enzyme and on its tryptic cleavage pattern. | |
| | |
MedLine Citation:
|
PMID: 2139606 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
The reversible inhibition of the sarcoplasmic-reticulum calcium-transport enzyme by pressure at room temperature is accompanied by a significant enhancement of the accessibility of the enzyme to tryptic cleavage dependent on the presence of calcium. The calcium-transport enzyme activity was monitored with dinitrophenyl phosphate as substrate. Pressure in the range 0.1-100.0 MPa affects trypsin cleavage of the control substrate N-alpha-benzoyl-L-arginine-4-nitroanilide hydrochloride little in the presence and absence of calcium. In contrast, application of 100.0 MPa to the calcium-transport enzyme at room temperature accelerates subsequent tryptic cleavage at the T2 but not at the T1 cleavage site [C. J. Brandl et al. (1986) Cell 44, 597-607]. Pressure application during tryptic digestion likewise solely affects cleavage at T2 which proceeds slowly in the absence but rapidly in the presence of calcium. At atmospheric pressure in the absence of calcium and at high pressure in the absence and presence of calcium new cleavage sites are exposed giving rise to new subfragments B1-3 in addition to the established peptides A1 and A2. Under pressure and in the presence of calcium, A1 and A2 rapidly disappear indicating the presence of calcium-binding sites in these peptides. In contrast, the B1-3 peptides which are most likely derivates of the B fragment accumulate in the presence and absence of calcium. In contrast to tryptic cleavage at atmospheric pressure, tryptic cleavage of the A as well as the B fragment tends to completion under pressure. In parallel to the disappearance of the A and B fragments calcium-dependent substrate hydrolysis vanishes. Computation of activation volumes for pressure-induced reversible enzyme inhibition and for tryptic cleavage furnished closely related volumes of opposite signs of 20-40 ml/mol and 80-100 ml/mol in the ranges 0.1-40.0 MPa and 40.0-100.0 MPa, respectively. Thus pressure produces reversible changes in the calcium-transport enzyme which activates and modifies tryptic-cleavage patterns at the T2 site of the A segment and at sites in its subfragments in the presence of calcium, i.e. if the enzyme residues in its E1 state. In contrast tryptic cleavage of the B fragment is accelerated by pressure independently of the presence of calcium. |
| | |
Authors:
|
N Ronzani; W Hasselbach; L Stephan |
Related Documents
:
|
20867586 - Structural transformations and anomalous viscosity in the b2o3 melt under high pressure. 19658966 - Pressure-induced invar behavior in pd3fe. 11715346 - High pressure effect on foaming properties of beta-lactoglobulin and dextran sulfate mi... 15697906 - Dynamics of the magnetic and structural alpha-epsilon phase transition in iron. 10701816 - Assessment of vascular aging and atherosclerosis in hypertensive subjects: second deriv... 11060186 - Influence of cholangiography on biliary sphincter of oddi manometric parameters. |
Publication Detail:
|
Type: Journal Article |
Journal Detail:
|
Title: European journal of biochemistry / FEBS Volume: 188 ISSN: 0014-2956 ISO Abbreviation: Eur. J. Biochem. Publication Date: 1990 Mar |
Date Detail:
|
Created Date: 1990-06-01 Completed Date: 1990-06-01 Revised Date: 2007-07-23 |
Medline Journal Info:
|
Nlm Unique ID: 0107600 Medline TA: Eur J Biochem Country: GERMANY, WEST |
Other Details:
|
Languages: eng Pagination: 557-65 Citation Subset: IM |
Affiliation:
|
Max-Planck-Institute für Medizinische Forschung, Abteilung für Physiologie, Heidelberg, Federal Republic of Germany. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Animals Annexin A6 Binding Sites Biological Transport Calcium / pharmacology* Calcium-Binding Proteins / isolation & purification Calcium-Transporting ATPases / antagonists & inhibitors, metabolism* Enzyme Activation Enzyme Stability Fluorescein-5-isothiocyanate Fluoresceins Maleimides Pressure Rabbits Sarcoplasmic Reticulum / enzymology* Temperature Thiocyanates Trypsin |
| Chemical | |
Reg. No./Substance:
|
0/Annexin A6; 0/Calcium-Binding Proteins; 0/Fluoresceins; 0/Maleimides; 0/Thiocyanates; 3326-32-7/Fluorescein-5-isothiocyanate; 55145-14-7/N-(7-dimethylamino-4-methylcoumarinyl)maleimide; 7440-70-2/Calcium; EC 3.4.21.4/Trypsin; EC 3.6.1.8/Calcium-Transporting ATPases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Phosphorylation of high-Mr caldesmon by protein kinase C modulates the regulatory function of this p...
Next Document: Purification and further characterization of the second nitrate reductase of Escherichia coli K12.