CaMKII uses GTP as a phosphate donor for both substrate and autophosphorylation |
| |
Authors: | S Lynn Bostrom Leslie C Griffith |
| |
Institution: | Department of Biology, National Center for Behavioral Genomics and Volen Center for Complex Systems, Brandeis University, 415 South St., Waltham, MA 02454-9110, United States |
| |
Abstract: | The vast majority of serine/threonine protein kinases have a strong preference for ATP over GTP as a phosphate donor. CK2 (Casein kinase 2) is an exception to this rule and in this study we investigate whether calcium/calmodulin-dependent protein kinase II (CaMKII) has the same extended nucleotide range. Using the Drosophila enzyme, we have shown that CaMKII uses Mg2+GTP with a higher Km and Vmax compared to Mg2+ATP. Substitution of Mn2+ for Mg2+ resulted in a much lower Km for GTP, while nearly abolishing the ability of CaMKII to use ATP. These similar results were obtained with rat αCaMKII, showing the ability to use GTP to be a general property of CaMKII. The Vmax difference between Mg2+ATP and Mg2+GTP was found to be due to the fact that ADP is a potent inhibitor of phosphorylation, while GDP has modest effects. There were no differences found between sites autophosphorylated by ATP and GTP, either by partial proteolysis or mass spectrometry. Phosphorylation of fly head extract revealed that similar proteins are substrates for CaMKII whether using Mg2+ATP or Mg2+GTP. This new information confirms that CaMKII can use both ATP and GTP, and opens new avenues for the study of regulation of this kinase. |
| |
Keywords: | Calcium/calmodulin-dependent protein kinase II (CaMKII) GTP Phosphorylation Kinetics CK2 (casein kinase II) Autophosphorylation |
本文献已被 ScienceDirect 等数据库收录! |
|