biostudies-literatureMayer LBritish Heart FoundationMedical Research CouncilNational Institute for Health Research (NIHR)RosetreesWellcome Trust1000-1011https://www.ebi.ac.uk/biostudies/studies/S-EPMC5877783biostudies-literatureUnknown131(9)Mutations in <i>NBEAL2</i>, the gene encoding the scaffolding protein Nbeal2, are causal of gray platelet syndrome (GPS), a rare recessive bleeding disorder characterized by platelets lacking α-granules and progressive marrow fibrosis. We present here the interactome of Nbeal2 with additional validation by reverse immunoprecipitation of Dock7, Sec16a, and Vac14 as interactors of Nbeal2. We show that GPS-causing mutations in its BEACH domain have profound and possible effects on the interaction with Dock7 and Vac14, respectively. Proximity ligation assays show that these 2 proteins are physically proximal to Nbeal2 in human megakaryocytes. In addition, we demonstrate that Nbeal2 is primarily localized in the cytoplasm and Dock7 on the membrane of or in α-granules. Interestingly, platelets fBloodNbeal2 interacts with Dock7, Sec16a, and Vac14.PMC5877783HDR-1004WT098503MR/L003120/1RG/13/13/30194MR/P02002X/1NF-SI-0513-10151NF-SI-0512-10165RE/13/6/30180M797HDR-9004106169/Z/14/ZNF-SI-0617-10113Mayer LYu LJasztal MYue WWBariana TKFavier RSmethurst PAGuerrero JAMeacham SAstle WJGrassi LPardo MAguera de Haro SChoudhary JOuwehand WHCollins JPetersen RNurden PfalseNbeal2 interacts with Dock7, Sec16a, and Vac14.Mutations in <i>NBEAL2</i>, the gene encoding the scaffolding protein Nbeal2, are causal of gray platelet syndrome (GPS), a rare recessive bleeding disorder characterized by platelets lacking α-granules and progressive marrow fibrosis. We present here the interactome of Nbeal2 with additional validation by reverse immunoprecipitation of Dock7, Sec16a, and Vac14 as interactors of Nbeal2. We show that GPS-causing mutations in its BEACH domain have profound and possible effects on the interaction with Dock7 and Vac14, respectively. Proximity ligation assays show that these 2 proteins are physically proximal to Nbeal2 in human megakaryocytes. In addition, we demonstrate that Nbeal2 is primarily localized in the cytoplasm and Dock7 on the membrane of or in α-granules. Interestingly, platelets f2018-01-01T00:00:00Z2018 Mar2026-04-30T18:11:41.276Z2019-03-26T23:52:42ZS-EPMC58777832918738010.1182/blood-2017-08-800359