CAP-50, a newly identified annexin, localizes in nuclei of cultured fibroblast 3Y1 cells

Akihiro Mizutani, Nobuteru Usuda, Hiroshi Tokumitsu, Hiroyuki Minami, Kiyoshi Yasui, Ryoji Kobayashi, Hiroyoshi Hidaka

Research output: Contribution to journalArticle

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Abstract

A 50-kDa protein, which binds to the growth-regulated gene (2A9) product, calcyclin in a calcium-dependent manner, was purified from bovine lung. Partial amino acid sequencing of the protein revealed it to be the bovine equivalent of rabbit lung CAP-50 (calcyclin-associated grotein, 50 kDa), which is a member of the annexin family and binds to calcyclin in a calcium-dependent manner. Specific polyclonal antibodies to bovine lung CAP-50 were prepared. Comparative studies between CAP-50 and synexin (annexin VII) on the immunoreactivity against anti-CAP-50 antibodies and the ability of binding to calcyclin revealed that CAP-50 was a distinct molecule from synexin. Using specific polyclonal antibodies to bovine lung CAP-50, tissue distribution and subcellular distribution of CAP-50 were investigated. In most rat tissues, except those in the central nervous systems and kidney, CAP-50 is expressed at a high or moderate level. Both studies by subcellular fractionation and by indirect immunofluorescence staining of the rat embryonic fibroblast cell line, 3Y1, revealed that CAP-50 mainly localized in nuclei. Moreover, between the cells at interphase and at mitotic phase, different distributions of CAP-50 were observed. That is, in the cells at interphase, CAP-50 seemed to localize throughout the nucleoplasm. On the other hand, in the cells during mitosis, CAP-50 was concentrated at the loop-like structure around the mitotic apparatus. CAP-50 was found in isolated 3Y1 nuclei lacking outer nuclear membranes, and approximately 50% of CAP-50 was extracted from the nuclei by chelating calcium. Thus, CAP-50, a unique annexin, localizes in nuclei.

Original languageEnglish
Pages (from-to)13498-13504
Number of pages7
JournalJournal of Biological Chemistry
Volume267
Issue number19
Publication statusPublished - Jul 5 1992
Externally publishedYes

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Annexins
Fibroblasts
Annexin A7
Lung
Interphase
calcyclin-associated protein 50
Calcium
Antibodies
Rats
Tissue
Spindle Apparatus
Aptitude
Protein Sequence Analysis
Nuclear Envelope
Neurology
Tissue Distribution
Indirect Fluorescent Antibody Technique
Fractionation
Chelation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mizutani, A., Usuda, N., Tokumitsu, H., Minami, H., Yasui, K., Kobayashi, R., & Hidaka, H. (1992). CAP-50, a newly identified annexin, localizes in nuclei of cultured fibroblast 3Y1 cells. Journal of Biological Chemistry, 267(19), 13498-13504.

CAP-50, a newly identified annexin, localizes in nuclei of cultured fibroblast 3Y1 cells. / Mizutani, Akihiro; Usuda, Nobuteru; Tokumitsu, Hiroshi; Minami, Hiroyuki; Yasui, Kiyoshi; Kobayashi, Ryoji; Hidaka, Hiroyoshi.

In: Journal of Biological Chemistry, Vol. 267, No. 19, 05.07.1992, p. 13498-13504.

Research output: Contribution to journalArticle

Mizutani, A, Usuda, N, Tokumitsu, H, Minami, H, Yasui, K, Kobayashi, R & Hidaka, H 1992, 'CAP-50, a newly identified annexin, localizes in nuclei of cultured fibroblast 3Y1 cells', Journal of Biological Chemistry, vol. 267, no. 19, pp. 13498-13504.
Mizutani A, Usuda N, Tokumitsu H, Minami H, Yasui K, Kobayashi R et al. CAP-50, a newly identified annexin, localizes in nuclei of cultured fibroblast 3Y1 cells. Journal of Biological Chemistry. 1992 Jul 5;267(19):13498-13504.
Mizutani, Akihiro ; Usuda, Nobuteru ; Tokumitsu, Hiroshi ; Minami, Hiroyuki ; Yasui, Kiyoshi ; Kobayashi, Ryoji ; Hidaka, Hiroyoshi. / CAP-50, a newly identified annexin, localizes in nuclei of cultured fibroblast 3Y1 cells. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 19. pp. 13498-13504.
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abstract = "A 50-kDa protein, which binds to the growth-regulated gene (2A9) product, calcyclin in a calcium-dependent manner, was purified from bovine lung. Partial amino acid sequencing of the protein revealed it to be the bovine equivalent of rabbit lung CAP-50 (calcyclin-associated grotein, 50 kDa), which is a member of the annexin family and binds to calcyclin in a calcium-dependent manner. Specific polyclonal antibodies to bovine lung CAP-50 were prepared. Comparative studies between CAP-50 and synexin (annexin VII) on the immunoreactivity against anti-CAP-50 antibodies and the ability of binding to calcyclin revealed that CAP-50 was a distinct molecule from synexin. Using specific polyclonal antibodies to bovine lung CAP-50, tissue distribution and subcellular distribution of CAP-50 were investigated. In most rat tissues, except those in the central nervous systems and kidney, CAP-50 is expressed at a high or moderate level. Both studies by subcellular fractionation and by indirect immunofluorescence staining of the rat embryonic fibroblast cell line, 3Y1, revealed that CAP-50 mainly localized in nuclei. Moreover, between the cells at interphase and at mitotic phase, different distributions of CAP-50 were observed. That is, in the cells at interphase, CAP-50 seemed to localize throughout the nucleoplasm. On the other hand, in the cells during mitosis, CAP-50 was concentrated at the loop-like structure around the mitotic apparatus. CAP-50 was found in isolated 3Y1 nuclei lacking outer nuclear membranes, and approximately 50{\%} of CAP-50 was extracted from the nuclei by chelating calcium. Thus, CAP-50, a unique annexin, localizes in nuclei.",
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AU - Mizutani, Akihiro

AU - Usuda, Nobuteru

AU - Tokumitsu, Hiroshi

AU - Minami, Hiroyuki

AU - Yasui, Kiyoshi

AU - Kobayashi, Ryoji

AU - Hidaka, Hiroyoshi

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