A new approach for the detection of multiple protein kinases using monoclonal antibodies directed to the highly conserved region of protein kinases. 2003

Isamu Kameshita, and Toshiyuki Tsuge, and Tomoko Kinashi, and Shun Kinoshita, and Noriyuki Sueyoshi, and Atsuhiko Ishida, and Shigeru Taketani, and Yasushi Shigeri, and Yoshiro Tatsu, and Noboru Yumoto, and Katsuichiro Okazaki
Department of Life Sciences, Kagawa University, Kagawa 761-0795, Japan. kamesita@ag.kagawa-u.ac.jp

To explore the protein kinase family enzymes expressed in cells, we attempted to generate antibodies that could detect a wide variety of protein kinases. For the production of such antibodies, synthetic peptides corresponding to amino acid sequences of a highly conserved subdomain (subdomain VIB) of the protein kinase family were used for immunization. Among the various peptide antigens, a peptide with 16 amino acids, CVVHRDLKPENLLLAS, effectively produced polyclonal antibodies with broad cross-reactivities to protein kinases. Two monoclonal antibodies, designated M8C and M1C, detected a variety of protein kinases such as calmodulin-dependent protein kinase II, calmodulin-dependent protein kinase IV, cAMP-dependent protein kinase, and mitogen-activated protein kinases, on Western blotting. The antibodies also immunoprecipitated various protein kinases in cell extracts. Furthermore, these antibodies could be used for detection of positive clones in the expression cloning of various protein kinases. Among 39 positive clones obtained from mouse brain cDNA library, 36 clones were identified as cDNA clones for various known and novel protein serine/threonine kinases, suggesting that the antibodies reacted highly specifically with various protein kinases. These results indicate that the present monoclonal antibodies directed to multiple protein kinases will be a powerful tool for the detection of a variety of known and novel protein kinases in cells.

UI MeSH Term Description Entries
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D011233 Precipitin Tests Serologic tests in which a positive reaction manifested by visible CHEMICAL PRECIPITATION occurs when a soluble ANTIGEN reacts with its precipitins, i.e., ANTIBODIES that can form a precipitate. Precipitin Test,Test, Precipitin,Tests, Precipitin
D011494 Protein Kinases A family of enzymes that catalyze the conversion of ATP and a protein to ADP and a phosphoprotein. Protein Kinase,Kinase, Protein,Kinases, Protein
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000911 Antibodies, Monoclonal Antibodies produced by a single clone of cells. Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D015153 Blotting, Western Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes. Immunoblotting, Western,Western Blotting,Western Immunoblotting,Blot, Western,Immunoblot, Western,Western Blot,Western Immunoblot,Blots, Western,Blottings, Western,Immunoblots, Western,Immunoblottings, Western,Western Blots,Western Blottings,Western Immunoblots,Western Immunoblottings
D017124 Conserved Sequence A sequence of amino acids in a polypeptide or of nucleotides in DNA or RNA that is similar across multiple species. A known set of conserved sequences is represented by a CONSENSUS SEQUENCE. AMINO ACID MOTIFS are often composed of conserved sequences. Conserved Sequences,Sequence, Conserved,Sequences, Conserved
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus

Related Publications

Isamu Kameshita, and Toshiyuki Tsuge, and Tomoko Kinashi, and Shun Kinoshita, and Noriyuki Sueyoshi, and Atsuhiko Ishida, and Shigeru Taketani, and Yasushi Shigeri, and Yoshiro Tatsu, and Noboru Yumoto, and Katsuichiro Okazaki
January 1991, Current studies in hematology and blood transfusion,
Isamu Kameshita, and Toshiyuki Tsuge, and Tomoko Kinashi, and Shun Kinoshita, and Noriyuki Sueyoshi, and Atsuhiko Ishida, and Shigeru Taketani, and Yasushi Shigeri, and Yoshiro Tatsu, and Noboru Yumoto, and Katsuichiro Okazaki
August 2003, European journal of clinical investigation,
Isamu Kameshita, and Toshiyuki Tsuge, and Tomoko Kinashi, and Shun Kinoshita, and Noriyuki Sueyoshi, and Atsuhiko Ishida, and Shigeru Taketani, and Yasushi Shigeri, and Yoshiro Tatsu, and Noboru Yumoto, and Katsuichiro Okazaki
June 2009, PloS one,
Isamu Kameshita, and Toshiyuki Tsuge, and Tomoko Kinashi, and Shun Kinoshita, and Noriyuki Sueyoshi, and Atsuhiko Ishida, and Shigeru Taketani, and Yasushi Shigeri, and Yoshiro Tatsu, and Noboru Yumoto, and Katsuichiro Okazaki
July 1984, Journal of virology,
Isamu Kameshita, and Toshiyuki Tsuge, and Tomoko Kinashi, and Shun Kinoshita, and Noriyuki Sueyoshi, and Atsuhiko Ishida, and Shigeru Taketani, and Yasushi Shigeri, and Yoshiro Tatsu, and Noboru Yumoto, and Katsuichiro Okazaki
December 1989, Biochemistry international,
Isamu Kameshita, and Toshiyuki Tsuge, and Tomoko Kinashi, and Shun Kinoshita, and Noriyuki Sueyoshi, and Atsuhiko Ishida, and Shigeru Taketani, and Yasushi Shigeri, and Yoshiro Tatsu, and Noboru Yumoto, and Katsuichiro Okazaki
January 1988, Kaku igaku. The Japanese journal of nuclear medicine,
Isamu Kameshita, and Toshiyuki Tsuge, and Tomoko Kinashi, and Shun Kinoshita, and Noriyuki Sueyoshi, and Atsuhiko Ishida, and Shigeru Taketani, and Yasushi Shigeri, and Yoshiro Tatsu, and Noboru Yumoto, and Katsuichiro Okazaki
July 2012, Biochemical and biophysical research communications,
Isamu Kameshita, and Toshiyuki Tsuge, and Tomoko Kinashi, and Shun Kinoshita, and Noriyuki Sueyoshi, and Atsuhiko Ishida, and Shigeru Taketani, and Yasushi Shigeri, and Yoshiro Tatsu, and Noboru Yumoto, and Katsuichiro Okazaki
January 2010, Bioscience, biotechnology, and biochemistry,
Isamu Kameshita, and Toshiyuki Tsuge, and Tomoko Kinashi, and Shun Kinoshita, and Noriyuki Sueyoshi, and Atsuhiko Ishida, and Shigeru Taketani, and Yasushi Shigeri, and Yoshiro Tatsu, and Noboru Yumoto, and Katsuichiro Okazaki
December 1992, Journal of immunological methods,
Isamu Kameshita, and Toshiyuki Tsuge, and Tomoko Kinashi, and Shun Kinoshita, and Noriyuki Sueyoshi, and Atsuhiko Ishida, and Shigeru Taketani, and Yasushi Shigeri, and Yoshiro Tatsu, and Noboru Yumoto, and Katsuichiro Okazaki
January 2008, Immunobiology,
Copied contents to your clipboard!