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

Huang,He; Ren,Hui-Min; Shang,Xi-Liang; Liu,Xue-Yuan

doi:10.3892/mmr.2014.2439

October 2014 Volume 10 Issue 4

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October 2014 Volume 10 Issue 4

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Article

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 / Copyright

Affiliations: Institute of Neurology, Fudan University, Shanghai 200040, P.R. China, Department of Sports Medicine, Fudan University, Shanghai 200040, P.R. China, Department of Neurology, Shanghai Tenth People's Hospital of Tongji University, Shanghai 200092, P.R. China
Pages: 1887-1892
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Published online on: August 4, 2014

https://doi.org/10.3892/mmr.2014.2439
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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.

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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

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

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