ERcalcistorin/protein disulfide isomerase (PDI). Sequence determination and expression of a cDNA clone encoding a calcium storage protein with PDI activity from endoplasmic reticulum of the sea urchin egg.
ERcalcistorin/protein disulfide isomerase (PDI). Sequence determination and expression of a cDNA clone encoding a calcium storage protein with PDI activity from endoplasmic reticulum of the sea urchin egg.
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发表时间：
1994-09
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通讯作者：
H. Lucero;D. Lebeche;B. Kaminer
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作者：
H. Lucero;D. Lebeche;B. Kaminer
在从海胆卵的内质网(ER)中纯化出一种58 kDa的类钙调素蛋白(Oberdorf，J.A.，Lebeche，D.，Head，J.F.和Kaminer，B.(1988)J.Biol.化学。263,6806-6809)，并鉴定其为高容量、低亲和力的钙结合蛋白(Lebeche，D.和Kaminer，B.(1992)Biochem)。J.287,741-747)，我们分离并测序了该蛋白的编码基因。推测的496个氨基酸含有一个17个残基的NH2端信号肽、一个KDEL COOH端ER保留信号和两个与蛋白质二硫键异构酶(PDI)相同的硫氧还蛋白活性部位结构域-CGHC-。海胆卵蛋白与哺乳动物PDI的序列同源性为55%，其PDI活性是兔肝PDI活性的30%。在海胆的卵母细胞、成熟卵、胚胎和分化组织中发现了不同数量的相应的mRNA。将该基因导入COS-7细胞，表达了一种能与抗海胆卵蛋白抗体发生免疫反应的58 kDa蛋白。这种分子似乎具有钙储存和内质网内PDI活性的双重功能。因此，我们将其重新命名为ERcalecstorin/PDI(ECAST/PDI)，这是一种有别于钙调素和钙网织蛋白的蛋白质。
Following the purification of a 58-kDa calsequestrin-like protein from the endoplasmic reticulum (ER) of sea urchin eggs (Oberdorf, J.A., Lebeche, D., Head, J. F., and Kaminer, B. (1988) J. Biol. Chem. 263, 6806-6809) and its characterization as a high capacity, low affinity calcium-binding protein (Lebeche, D., and Kaminer, B. (1992) Biochem. J. 287, 741-747) we isolated and sequenced a cDNA encoding for this protein. The deduced 496 amino acids contain a 17-residue NH2-terminal signal peptide, a KDEL COOH-terminal ER retention signal and two thioredoxin-like active site domains, -CGHC-, identical with those in protein disulfide isomerase (PDI). The sea urchin egg protein shares a 55% sequence identity with mammalian PDI and its PDI activity is 30% of the activity of rabbit liver PDI. The corresponding mRNA was found in oocytes, mature eggs, embryos, and differentiated tissues of the sea urchin in varying amounts. COS-7 cells transfected with the cDNA, expressed a 58-kDa protein immunoreactive to antibodies against the sea urchin egg protein. This molecule appears to have a dual function of calcium storage and PDI activity within the ER. We hence redesignate it ERcalcistorin/PDI (ECaSt/PDI), a protein that is distinct from calsequestrin and calreticulin.