Title A Modified Immunoblot Method to Identify Substrates of Protein Kinases
Author Choong-Min Kang1*, Wan Jin Jahng2, Robert N. Husson3, and Sang Hee Lee4*
Address 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 Korea
Bibliography Journal of Microbiology, 49(3),499-501, 2011,
Key Words modified immunoblot method, substrates of kinases, partial transfer of proteins
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.