Biopolym. Cell. 2014; 30(1):74-76.
Short Communications
Ca/calmodulin-dependent phosphorylation of endocytic scaffold ITSN1
1Morderer D. Ye., 1Nikolaienko O. V., 1Skrypkina I. Ya., 1Rymarenko O. V., 1Kropyvko S. V., 1Tsyba L. O., 1Rynditch A. V.
  1. State Key Laboratory of Molecular and Cellular Biology
    Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680


ITSN1 is an endocytic scaffold protein with a prominent function in synaptic transmission. It is known that Ca signaling is crucial for the regulation of synaptic proteins functioning. Aim. Checking the possibility of Ca/calmodulin-dependent phosphorylation of ITSN1. Methods. Affinity chromatography, in vitro kinase reaction, Western blotting, gel staining with fluorescent stains. Results. We show that the fraction of calmodulin-binding proteins is able to phosphorylate the recombinant fragments encoding the coiled-coil region and the SH3 domain-containing region of ITSN1 in the presence of Ca ions and calmodulin. Conclusions. The coiled-coil region and the SH3 domain-containing region of ITSN1 undergo Ca/calmodulin-dependent phosphorylation in vitro, suggesting a possible regulation of ITSN1 by Ca signaling.
Keywords: ITSN1, phosphorylation, calmodulin, Ca signaling


[1] Tsyba L., Nikolaienko O., Dergai O., Dergai M., Novokhatska O., Skrypkina I., Rynditch A. Intersectin multidomain adaptor proteins: regulation of functional diversity Gene 2011 473, N 2:67–75.
[2] O'Bryan J. P. Intersecting pathways in cell biology Sci. Signal 2010 3, N 152:re10.
[3] Tsyba L. O., Dergai M. V., Skrypkina I. Ya., Nikolaienko O. V., Dergai O. V., Kropyvko S. V., Novokhatska O. V., Morderer D. Ye., Gryaznova T. A., Gubar O. S., Rynditch A. V. ITSN protein family: regulation of diversity, role in signalling and pathology. Biopolym. Cell. 2013; 29, N 3:244–51.
[4] Ballif B. A., Villen J., Beausoleil S. A., Schwartz D., Gygi S. P. Phosphoproteomic analysis of the developing mouse brain Mol. Cell Proteomics 2004 3, N 11:1093–101.
[5] Dergai O., Dergai M., Skrypkina I., Matskova L., Tsyba L., Gudkova D., Rynditch A. The LMP2A protein of Epstein-Barr virus regulates phosphorylation of ITSN1 and Shb adaptors by tyrosine kinases Cell. Signal 2013 25, N 1:33–40.
[6] Pechstein A., Shupliakov O., Haucke V. Intersectin 1: a versatile actor in the synaptic vesicle cycle Biochem. Soc. Trans 2010 38, Pt 1:181–6.
[7] Kostyuk P. G. Key role of calcium signaling in synaptic transmission Neurophysiology 2007 39, N 4–5:248–50.
[8] Evergren E., Gad H., Walther K., Sundborger A., Tomilin N., Shupliakov O. Intersectin is a negative regulator of dynamin recruitment to the synaptic endocytic zone in the central synapse J. Neurosci 2007 27, N 2:379–90.
[9] Winther A. M., Jiao W, Vorontsova O., Rees K. A., Koh T. W., Sopova E., Schulze K. L., Bellen H. J., Shupliakov O. The dynamin-binding domains of Dap160/intersectin affect bulk membrane retrieval in synapses J. Cell. Sci 2013 126, Pt 4:1021–31.
[10] Nishimura T., Yamaguchi T., Tokunaga A., Hara A., Hamaguchi T., Kato K., Iwamatsu A., Okano H., Kaibuchi K. Role of numb in dendritic spine development with a Cdc42 GEF intersectin and EphB2 Mol. Biol. Cell 2006 17, N 3:1273–85.
[11] Wayman G. A., Lee Y. S., Tokumitsu H., Silva A. J., Soderling T. R. Calmodulin-kinases: modulators of neuronal development and plasticity Neuron 2008 59, N 6:914–31.