Ca/calmodulin-dependent phosphorylation of endocytic scaffold ITSN1

Morderer, D. Ye.; Nikolaienko, O. V.; Skrypkina, I. Ya.; Rymarenko, O. V.; Kropyvko, S. V.; Tsyba, L. O.; Rynditch, A. V.

doi:10.7124/bc.000882

Biopolym. Cell. 2014; 30(1):74-76.

http://dx.doi.org/10.7124/bc.000882

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.

State Key Laboratory of Molecular and Cellular Biology
Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680

Abstract

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

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