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Research Support, Non-U.S. Gov't
- A Modified Immunoblot Method to Identify Substrates of Protein Kinases
- Choong-Min Kang 1, Wan Jin Jahng 2, Robert N. Husson 3, Sang Hee Lee 4
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Journal of Microbiology 2011;49(3):499-501
DOI: https://doi.org/10.1007/s12275-011-0465-y
Published online: June 30, 2011
1Department of Biological Science, Wayne State University, 5047 Gullen Mall, Detroit, MI 48202, USA, 2Department of Biological Sciences, Michigan Technological University, Houghton, MI 49931, USA, 3Division of Infectious Diseases, Children’s Hospital Boston, Harvard Medical School, 300 Longwood Ave., Boston, MA 02115, USA, 4Department of Biological Sciences, Myongji University, Gyeonggido 449-728, Republic of Korea1Department of Biological Science, Wayne State University, 5047 Gullen Mall, Detroit, MI 48202, USA, 2Department of Biological Sciences, Michigan Technological University, Houghton, MI 49931, USA, 3Division of Infectious Diseases, Children’s Hospital Boston, Harvard Medical School, 300 Longwood Ave., Boston, MA 02115, USA, 4Department of Biological Sciences, Myongji University, Gyeonggido 449-728, Republic of Korea
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Corresponding author:
Choong-Min Kang , Tel: +82-31-330-6195;,
Sang Hee Lee , Tel: +82-31-330-6195;,
Received: 16 November 2010 • Accepted: 13 January 2011
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Abstract
While protein kinases are key components in multiple cellular processes, efficient identification of cognate in vivo substrates remains challenging. Here we describe a powerful method to screen potential substrates of protein kinases by partial transfer of proteins from a 2D-PAGE gel to a Western blot membrane. This approach allowed precise pinpointing of candidate substrate spots in the 2D gel, and identifying physiological substrates of protein kinases in Mycobacterium tuberculosis.
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