Receptor-like protein tyrosine phosphatase alpha (RPTP alpha or PTP alpha), a type I transmembrane glycoprotein with complex N-glycans, executes multifunction roles on cell behaviors. However, its effect on tumorigenesis and metastasis remains controversial. In this study, PTP alpha is identified as a novel substrate of N-Acetylglucosaminyltransferase V (GnT-V). Immunofluorescence results showed that addition of beta 1,6 GlcNAc branches on PTP alpha enhanced PTP alpha's cytomembrane assemble in GnT-V-MCF-7 compared with Mock-MCF-7 (MCF7 cells transfected with the vector pcDNA3). Then we found the alleviating degradation of PTP alpha was observed in GnT-V-MCF-7 while PTP alpha in Mock-MCF-7 was prone to quick degradation. Increased cell-surface retention subsequently enhanced PTP alpha's catalytic activity on the dephosphorylation of Src kinase at Tyr529 and promoted focal adhesion formation and mature. Therefore, our findings could provide an insight into the molecular mechanism of how GnT-V promoted cell migration, suggesting that PTP alpha could be one of factors regulating promote migration of breast cancer cell. (C) 2016 Elsevier Inc. All rights reserved.