Molecular dynamics in mouse atrial tumor sarcoplasmic reticulum.

Abstract

We have determined directly the effects of the inhibitory peptide phospholamban (PLB) on the rotational dynamics of the calcium pump (Ca-ATPase) of cardiac sarcoplasmic reticulum (SR). This was accomplished by comparing mouse ventricular SR, which has PLB levels similar to those found in other mammals, with mouse atrial SR, which is effectively devoid of PLB and thus has much higher (unregulated) calcium pump activity. To obtain sufficient quantities of atrial SR, we isolated the membranes from atrial tumor cells. We used time-resolved phosphorescence anisotropy of an erythrosin isothiocyanate label attached selectively and rigidly to the Ca-ATPase, to detect the microsecond rotational motion of the Ca-ATPase in the two preparations. The time-resolved phosphorescence anisotropy decays of both preparations at 25 degrees C were multi-exponential, because of the presence of different oligomeric species. The rotational correlation times for the different oligomers were similar for the two preparations, but the total decay amplitude was substantially greater for atrial tumor SR, indicating that a smaller fraction of the Ca-ATPase molecules exists as large aggregates. Phosphorylation of PLB in ventricular SR decreased the population of large-scale Ca-ATPase aggregates to a level similar to that of atrial tumor SR. Lipid chain mobility (fluidity), detected by electron paramagnetic resonance of stearic acid spin labels, was very similar in the two preparations, indicating that the higher protein mobility in atrial tumor SR is not due to higher lipid fluidity. We conclude that PLB inhibits by inducing Ca-ATPase lateral aggregation, which can be relieved either by phosphorylating or removing PLB.

• Authors: J C Voss, J E Mahaney, L R Jones, D D Thomas
• Journal: Biophysical journal (Biophys J) Vol. 68 Issue 5 Pg. 1787-95 (May 1995) ISSN: 0006-3495 [Print] United States
• PMID: 7612820 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Calcium-Binding Proteins
• Lipid Bilayers
• phospholamban
• Calcium-Transporting ATPases

Topics

• Animals
• Calcium-Binding Proteins (metabolism, pharmacology)
• Calcium-Transporting ATPases (chemistry, isolation & purification, metabolism)
• Electrophoresis, Polyacrylamide Gel
• Fluorescence Polarization
• Heart Atria
• Heart Neoplasms (enzymology)
• Heart Ventricles
• Kinetics
• Lipid Bilayers
• Luminescence
• Mammals
• Membrane Fluidity
• Mice
• Models, Structural
• Myocardium (enzymology)
• Phosphorylation
• Protein Conformation
• Rotation
• Sarcoplasmic Reticulum (enzymology, ultrastructure)