Detection of the phosphatase activity of carbonic anhydrase III on a nitrocellulose membrane following 2D gel electrophoresis

  • Authors:
    • He Huang
    • Hui-Min Ren
    • Xi-Liang Shang
    • Xue-Yuan Liu
  • View Affiliations

  • Published online on: August 4, 2014     https://doi.org/10.3892/mmr.2014.2439
  • Pages: 1887-1892
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Carbonic anhydrase isozyme III (CAIII) is unique among the carbonic anhydrases because it exhibits phosphatase activity. CAIII is relatively specific to skeletal muscles, and may therefore be a useful diagnostic marker for muscular diseases. In the muscles of patients with myasthenia gravis (MG), CAIII is deficient and previous studies have demonstrated that changes in the phosphatase activity of CAIII is a fundamental mechanism underlying the weakness and fatigability of MG. However, there have been no effective analytical methods for investigating its phosphatase activity until now. In the present study, a new method combining two-dimensional electrophoresis (2-DE) and phosphatase staining in situ on a nitrocellulose membrane was reported to detect the phosphatase of CAIII in skeletal muscle extracts. Furthermore, a recombinant CAIII was constructed and its phosphatase activity staining was demonstrated to be positive. This method allows for the effective detection of the phosphatase activity of CAIII following 2-DE and is a promising technique for functional proteomics.
View Figures
View References

Related Articles

Journal Cover

October 2014
Volume 10 Issue 4

Print ISSN: 1791-2997
Online ISSN:1791-3004

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Huang H, Ren H, Shang X and Liu X: Detection of the phosphatase activity of carbonic anhydrase III on a nitrocellulose membrane following 2D gel electrophoresis. Mol Med Rep 10: 1887-1892, 2014.
APA
Huang, H., Ren, H., Shang, X., & Liu, X. (2014). Detection of the phosphatase activity of carbonic anhydrase III on a nitrocellulose membrane following 2D gel electrophoresis. Molecular Medicine Reports, 10, 1887-1892. https://doi.org/10.3892/mmr.2014.2439
MLA
Huang, H., Ren, H., Shang, X., Liu, X."Detection of the phosphatase activity of carbonic anhydrase III on a nitrocellulose membrane following 2D gel electrophoresis". Molecular Medicine Reports 10.4 (2014): 1887-1892.
Chicago
Huang, H., Ren, H., Shang, X., Liu, X."Detection of the phosphatase activity of carbonic anhydrase III on a nitrocellulose membrane following 2D gel electrophoresis". Molecular Medicine Reports 10, no. 4 (2014): 1887-1892. https://doi.org/10.3892/mmr.2014.2439