Lockyer P J, Puente E, Windass J, Earley F, East J M, Lee A G
Division of Biochemistry and Molecular Biology, School of Biological Sciences, University of Southampton, UK.
Biochim Biophys Acta. 1998 Feb 2;1369(1):14-8. doi: 10.1016/s0005-2736(97)00247-2.
A complementary DNA for the Tobacco Budworm, Heliothis virescens, sarco(endo)plasmic reticulum-type Ca(2+)-ATPase (HVSERCA) has been cloned and sequenced. cDNA fragments of adult rabbit fast-twitch muscle Ca(2+)-ATPase (SERCA1a) were used as heterologous probes to isolate a partial cDNA clone coding for a protein with high homology to the Ca(2+)-ATPase from Drosophila melanogaster (DRSERCA) and vertebrate ER/SR Ca2+ pumps. The entire cDNA clone contains an ORF encoding a protein of 1000 amino acids which shares the characteristic motifs of a P-type ATPase. HVSERCA shares 89% identity with DRSERCA, 80% identity with the Artemia Ca(2+)-ATPase and 72% identity with avian and mammalian SERCAs. An insect Ca(2+)-ATPase-specific polyclonal antiserum has been raised against a fusion protein containing sequence from the cytoplasmic domain of HVSERCA. Heterologous expression of the insect pump in COS-7 cells has been demonstrated by immunocytochemistry and the reticular pattern of staining is consistent with an ER localisation. However, the expressed enzyme from COS-7 cells does not appear to be active.
烟草夜蛾（Heliothis virescens）肌质（内质）网型Ca(2+)-ATP酶（HVSERCA）的互补DNA已被克隆和测序。成年兔快肌Ca(2+)-ATP酶（SERCA1a）的cDNA片段被用作异源探针，以分离一个部分cDNA克隆，该克隆编码一种与黑腹果蝇（Drosophila melanogaster，DRSERCA）的Ca(2+)-ATP酶以及脊椎动物内质网/肌质网Ca2+泵具有高度同源性的蛋白质。整个cDNA克隆包含一个编码1000个氨基酸的蛋白质的开放阅读框，该蛋白质具有P型ATP酶的特征基序。HVSERCA与DRSERCA的同一性为89%，与卤虫Ca(2+)-ATP酶的同一性为80%，与鸟类和哺乳动物的SERCA的同一性为72%。已经制备了一种针对含有HVSERCA胞质结构域序列的融合蛋白的昆虫Ca(2+)-ATP酶特异性多克隆抗血清。通过免疫细胞化学证明了昆虫泵在COS-7细胞中的异源表达，染色的网状模式与内质网定位一致。然而，COS-7细胞中表达的酶似乎没有活性。
