%0 Generic %A A., Hafner %A N., Obermajer %A J., Kos %D 2011 %T Supplementary Material for: Gamma-1-Syntrophin Mediates Trafficking of Gamma-Enolase towards the Plasma Membrane and Enhances Its Neurotrophic Activity %U https://karger.figshare.com/articles/dataset/Supplementary_Material_for_Gamma-1-Syntrophin_Mediates_Trafficking_of_Gamma-Enolase_towards_the_Plasma_Membrane_and_Enhances_Its_Neurotrophic_Activity/5121712 %R 10.6084/m9.figshare.5121712.v1 %2 https://ndownloader.figshare.com/files/8706364 %K Gamma-1-syntrophin %K Gamma-enolase %K Cathepsin X %K Intracellular trafficking %K Neurotrophic effect %X Syntrophins are scaffold proteins that can bind several signaling molecules and localize them to the plasma membrane. We demonstrate here that in neuroblastoma SH-SY5Y cells, brain-specific γ1-syntrophin binds the neurotrophic factor γ-enolase through its PDZ domain, and translocates it to the plasma membrane, as shown by immunoprecipitation, surface plasmon resonance, fluorescence colocalization and flow cytometry. Extensive colocalization of γ1-syntrophin and γ-enolase was observed in neurite growth cones in differentiated SH-SY5Y cells. Silencing of the γ1-syntrophin gene by RNA interference significantly reduced the re-distribution of γ-enolase to the plasma membrane and impaired its neurotrophic effects. We demonstrated that an intact C-terminal end of γ-enolase is essential for its γ1-syntrophin-assisted trafficking. The cleavage of two amino acids at the C-terminal end of γ-enolase by the carboxypeptidase cathepsin X prevents binding with the γ1-syntrophin PDZ domain. Collectively, these data demonstrate that γ1-syntrophin participates in γ-enolase translocation towards the plasma membrane, a pre-requisite for its neurotrophic activity. By disrupting this γ1-syntrophin-guided subcellular distribution, cathepsin X reduces γ-enolase-induced neurotrophic signaling. %I Karger Publishers