Annexin V is localized in association with Z-line of rat cardiac myocytes

Luming Wang, Md Mohibur Rahman, Hiroshi Iida, Tetsuichiro Inai, Shun-Ichiro Kawabata, Sadaaki Iwanaga, Yosaburo Shibata

Research output: Contribution to journalArticle

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Abstract

Objective: The aim of this study was to characterize a 33-kDa protein (p33) isolated from bovine liver and to determine the subcellular localization of the protein in rat cardiocytes as well as in non-cardiac tissues. Methods: Cycles of calcium-induced precipitation coupled with EGTA-resolubilization were used to isolate crude annexins from bovine and rat tissues. Column chromatography was performed to purify the p33 from the crude annexins. The protein was identified as annexin V by partial amino acid sequence determination. Specificity of anti-annexin V antibody was examined by using Western blotting after one- or two-dimensional electrophoresis. For characterization of the protein, immunofluorescence microscopy and actin-binding assays were carried out. Results: Immunofluorescence microscopy showed that annexin V was stained as a striated pattern along myofibrils on frozen sections of both atria and ventricles of adult rats. The striations were seen more clearly in cultured rat atrial myocytes. Examination of doubly stained cardiocytes with anti-annexin V and anti-α-actinin by a confocal laser scanning microscope suggests that annexin V is localized in association with the Z-line of rat cardiac myocytes. We also found that annexin V was stained intensely in actin-rich regions of non-cardiac tissues such as bile canaliculi of rat liver and brush border-terminal web region in the epithelial cells of both small intestine and kidney proximal tubular cells. F-actin binding experiments revealed that annexin V failed to bind to F-actin directly in vitro. Conclusion: Our results suggest that annexin V is a new component of the Z-line in rat cardiocytes and might be involved in regulation of its organization.

Original languageEnglish
Pages (from-to)363-371
Number of pages9
JournalCardiovascular research
Volume30
Issue number3
DOIs
Publication statusPublished - Jan 1 1995

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Annexin A5
Cardiac Myocytes
Actins
Annexins
Fluorescence Microscopy
Proteins
Bile Canaliculi
Actinin
Myofibrils
Liver
Egtazic Acid
Protein Sequence Analysis
Frozen Sections
Microvilli
Muscle Cells
Small Intestine
Electrophoresis
Chromatography
Lasers
Western Blotting

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

Annexin V is localized in association with Z-line of rat cardiac myocytes. / Wang, Luming; Rahman, Md Mohibur; Iida, Hiroshi; Inai, Tetsuichiro; Kawabata, Shun-Ichiro; Iwanaga, Sadaaki; Shibata, Yosaburo.

In: Cardiovascular research, Vol. 30, No. 3, 01.01.1995, p. 363-371.

Research output: Contribution to journalArticle

Wang, Luming ; Rahman, Md Mohibur ; Iida, Hiroshi ; Inai, Tetsuichiro ; Kawabata, Shun-Ichiro ; Iwanaga, Sadaaki ; Shibata, Yosaburo. / Annexin V is localized in association with Z-line of rat cardiac myocytes. In: Cardiovascular research. 1995 ; Vol. 30, No. 3. pp. 363-371.
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