DEMONSTRATION OF 2 ISOFORMS OF THE SERCA-2B TYPE CA2+,MG2+-ATPASE IN PANCREATIC ENDOPLASMIC-RETICULUM
DEMONSTRATION OF 2 ISOFORMS OF THE SERCA-2B TYPE CA2+,MG2+-ATPASE IN PANCREATIC ENDOPLASMIC-RETICULUM
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DOI：
10.1016/0005-2736(93)90253-v
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发表时间：
1993-11-07
期刊：
影响因子：
--
通讯作者：
WEBB, R
中科院分区：
文献类型：
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作者：
DORMER, RL;CAPURRO, DE;WEBB, R
用与SERCA-2b钙镁三磷酸腺苷酶C端序列相对应的12个氨基酸多肽的抗体从胰腺粗面内质网中沉淀出钙镁三磷酸腺苷酶活性。Thapsigargin和Vvadate对该活性的抑制作用与天然ER膜相同，且呈浓度依赖关系。用活性染料-琼脂糖亲和层析部分纯化了Ca~(2+)，Mg~(2+)-ATPase，激活了该酶，表明存在内源抑制物，该抑制物通过与活性染料结合而分离。免疫印迹和免疫沉淀蛋白分析显示，其分子质量约为两条带。111 kDa和97 kDa。结论：胰腺内质网钙镁ATPase为SERCA-2b型，由两种亚型组成。
An antibody raised against a 12 amino acid peptide corresponding to the C-terminal sequence of the SERCA-2b Ca2+,Mg2+-ATPase precipitated Ca2+,Mg2+-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+,Mg2+-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+,Mg2+-ATPase is of the SERCA-2b type and consists of two isoforms.