GTPase-activating protein oligophrenin 1 is a new partner of multifunctional adapter protein intersectin 1

• DOI: 10.7124/bc.000070
• Publication Date: 2012-11-25T10:45:04Z
• Authors: O. S. Gubar; S. Houy; P. Billuart; S. V. Kropyvko; L. O. Tsyba; S. Gasman; A. V. Rynditch
• Keywords: 0301 basic medicine; 0303 health sciences; 03 medical and health sciences; [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology; oligophrenin 1; intersectin 1; PC12; exocytosis

Intersectin 1 (ITSN1) is a multifunctional adaptor protein which is involved in endocytosis, exocytosis and cellular signaling and it is also associated with such pathologies as Down syndrome and Alzheimer’s disease. The aim of this study was to identify new ITSN1 protein partners which are implicated in membrane trafficking. Methods. In silico analysis by Scansite online resource had identified a GTPase activating protein oligophrenin 1 (OPHN1) as a potential partner of ITSN1 SH3A domain. GST pull-down and immunoprecipitation were used to prove complex formation between ITSN1 and OPHN1. Subcellular protein localization was determined by immunofluorescence and confocal microscopy. Results. We have shown that brain-specific and ubiquitously expressed SH3A domain isoforms of ITSN1 interact with OPHN1. ITSN1 and OPHN1 form complexes in both resting and stimulated to exocytosis PC12 cell line. Conclusions. GTPase activating protein OPHN1 and adaptor protein ITSN1 interact in PC12 cell line independently of exocytosis stimulation.