Calreticulin-2 is localized in the lumen of the endoplasmic reticulum but is not a Ca2+-binding protein

Ryuji Nomura, Minami Orii, Takao Senda

研究成果: Article査読

4 被引用数 (Scopus)

抄録

Calreticulin (CRT)-1 is a major Ca2+-buffering protein in the lumen of the endoplasmic reticulum. Human and murine CRT-2 was isolated in 2002, but the subcellular localization and function is still unclear. Here, we studied the intracellular localization and function of CRT-2 with hemagglutinin-tagged (HA-) human CRT-2. Western blotting revealed HA-CRT-2 as a single band at 50 kDa. Using immunofluorescence microscopy of cultured fibroblasts and epithelial cells transfected with HA-CRT-2 cDNA, labeling for HA-CRT-2 was seen as a reticular network with a nuclear envelope pattern that colocalized with calnexin and protein disulfide isomerase. Immunoelectron microscopy confirmed that HA-CRT-2 was localized in the lumen of the endoplasmic reticulum. Stains-all staining, a method to detect Ca2+-binding proteins, could not stain the immunoprecipitate of HA-CRT-2, although HA-CRT-1 immunoprecipitate was stained blue. These results indicate that the molecular weight of the non-tagged CRT-2 on SDS-PAGE is 49 kDa, and that CRT-2, as well as CRT-1, is localized in the lumen of the endoplasmic reticulum, but that CRT-2 capacity for Ca2+-binding may be absent or much lower than that of CRT-1.

本文言語English
ページ(範囲)531-538
ページ数8
ジャーナルHistochemistry and Cell Biology
135
6
DOI
出版ステータスPublished - 06-2011

All Science Journal Classification (ASJC) codes

  • 組織学
  • 分子生物学
  • 医療検査技術
  • 細胞生物学

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