Differences in calcium uptake in native canine cardiac microsomes are correlated with the ratio of unphosphorylated to phosphorylated phospholamban as determined by Western blot analysis.
Phospholamban (PLM) is detectable by Western blot analysis of canine cardiac microsomes using rabbit antiserum against a peptide containing the 2 to 30 amino acid sequence of PLM. Phosphorylated PLM is distinguishable from the unphosphorylated form by virtue of a reduced electrophoretic mobility. Utilizing digital image analysis to determine relative band densities, it was found that the ratio of unphosphorylated to phosphorylated PLM is correlated with the rate of calcium uptake in 5 preparations of native microsomes (r = 0.94, p less than 0.01). The present analysis may be useful for determining the phosphorylation state of PLM in microsomes obtained from animals in physiological states characterized by impaired sarcoplasmic reticulum calcium pump activity.
Kasinathan
C
C
Department of Physiology and Biophysics, Mount Sinai School of Medicine, City University of New York, New York 10029.
Xu
Z C
ZC
Kirchberger
M A
MA
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002147:Calmodulin / Q000494:pharmacology; D000242:Cyclic AMP / Q000494:pharmacology; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D008861:Microsomes / Q000378:metabolism*; D008970:Molecular Weight; D009206:Myocardium / Q000378:metabolism*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D000099211:Phospholamban
D000255:Adenosine Triphosphate; D002118:Calcium; D002135:Calcium-Binding Proteins; D002147:Calmodulin; D000242:Cyclic AMP; D011494:Protein Kinases; D000099211:Phospholamban