Author(s)

Ellen Dirkx, Freek G. Bouwman, Didier Vertommen, Edwin C. Mariman, Sakthivel Sadayappan, Jan F. C. Glatz, Joost J. F. P. Luiken, Guillaume J. van Eys

Department of Cell and Molecular Physiology, Stritch School of Medicine, Loyola University, Chicago, USA.
Department of Human Biology, Nutrition and Toxicology Research Institute Maastricht (NUTRIM), Maastricht University, Maastricht, The Netherlands.
Department of Molecular Genetics, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, The Netherlands.
Protein Phosphorylation Unit, Université Catholique de Louvain, Brussels, Belgium.

Protein phosphorylation plays an important role in physiological processes, such as muscle contraction. Phospho-specific antibodies have become powerful tools to study these processes. Cardiac myosin binding protein-C (cMyBP-C) is one of the proteins that make up the contractile apparatus of cardiomyocytes. Phosphorylation of cMyBP-C is essential for normal cardiac function, since dephosphorylation of this protein leads to its degradation and has been associated with cardiomyopathy. One of the upstream kinases, which phosphorylate cMyBP-C, is protein kinase D (PKD). While studying the role of PKD in cMyBP-C phosphorylation, we tried to analyze phosphorylation of PKD with a phospho-specific PKD-Ser744/748 antibody. Contrary to the expected 115 kDa, a signal was found for a 150-kDa protein. By MALDI-TOF mass spectrometry, we identified this protein to be cMyBP-C. These data were confirmed by immunostaining using the p-PKD-Ser744/748 antibody, which displayed a striated pattern similar to the one observed for a regular cMyBP-C antibody. To our knowledge there are no antibodies commercially available for phosphorylated cMyBP-C. Thus, the p-PKD-Ser744/748 antibody can accelerate research into the role of cMyBP-C phosphorylation in cardiomyocytes.

Protein Kinase D; Cardiac Myosin Binding Protein-C; Phospho-Specific Antibody; Protein Phosphorylation

Dirkx, E. , Bouwman, F. , Vertommen, D. , Mariman, E. , Sadayappan, S. , Glatz, J. , Luiken, J. and Eys, G. (2013) Detection of cardiac myosin binding protein-C (cMyBP-C) by a phospho-specific PKD antibody in contracting rat cardiomyocytes. Advances in Bioscience and Biotechnology, 4, 1-6. doi: 10.4236/abb.2013.44A001.