Dormer R L, Capurro D E, Morris R, Webb R
Department of Medical Biochemistry, University of Wales College of Medicine, Heath Park, Cardiff, UK.
Biochim Biophys Acta. 1993 Nov 7;1152(2):225-30. doi: 10.1016/0005-2736(93)90253-v.
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.
一种针对与SERCA-2b Ca2+、Mg(2+)-ATP酶C末端序列相对应的12个氨基酸肽产生的抗体，从胰腺粗面内质网中沉淀出Ca2+、Mg(2+)-ATP酶活性。毒胡萝卜素和钒酸盐以与天然内质网膜相同的浓度依赖性抑制该活性。使用反应染料琼脂糖亲和色谱法对Ca2+、Mg(2+)-ATP酶进行部分纯化导致该酶激活，这表明存在一种内源性抑制剂，其通过与反应染料结合而被去除。免疫印迹和免疫沉淀蛋白分析显示出两条分子量约为111 kDa和97 kDa的条带。得出的结论是，胰腺内质网Ca2+、Mg(2+)-ATP酶属于SERCA-2b类型，由两种同工型组成。
