Demonstration of two isoforms of the SERCA-2b type Ca2+,Mg(2+)-ATPase in pancreatic endoplasmic reticulum.
An antibody raised against a 12 amino acid peptide corresponding to the C-terminal sequence of the SERCA-2b Ca2+,Mg(2+)-ATPase precipitated Ca2+,Mg(2+)-ATPase activity from pancreatic rough ER. Thapsigargin and vanadate inhibited the activity with the same concentration-dependence as for native ER membranes. Partial purification of Ca2+,Mg(2+)-ATPase using Reactive Dye-agarose affinity chromatography resulted in activation of the enzyme, suggesting the presence of an endogenous inhibitor which was detached by binding to the Reactive Dye. Immunoblots and analysis of immunoprecipitated protein revealed two bands of molecular masses approx. 111 kDa and 97 kDa. It is concluded that pancreatic ER Ca2+,Mg(2+)-ATPase is of the SERCA-2b type and consists of two isoforms.
Dormer
R L
RL
Department of Medical Biochemistry, University of Wales College of Medicine, Heath Park, Cardiff, UK.
Capurro
D E
DE
Morris
R
R
Webb
R
R
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D017301:Ca(2+) Mg(2+)-ATPase / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D003429:Cross Reactions; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D015151:Immunoblotting; D007527:Isoenzymes / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology*; D011233:Precipitin Tests; D051381:Rats; D017208:Rats, Wistar; D013729:Terpenes / Q000494:pharmacology; D019284:Thapsigargin; D014638:Vanadates / Q000494:pharmacology
D007527:Isoenzymes; D013729:Terpenes; D014638:Vanadates; D019284:Thapsigargin; D017301:Ca(2+) Mg(2+)-ATPase