NOK is a newly identified receptor protein-tyrosine kinase (PRTK) molecule that can promote tumorigenesis and metastasis. Previous data showed that NOK could activate the phosphatidylinositol 3'-kinase (PI3K) pathway in stable BaF3 cells. But how does NOK activate PI3K in cells remains unknown. It was showed that NOK physically interacted with the PI3K downstream effector Akt and enhanced its activation in human embryo kidney 293T (HEK293T) cells. Deletion mapping indicated that protein kinase B (Akt) was able to directly contact with the kinase domain of NOK. Inactivating the Akt kinase domain significantly reduced the intermolecular interaction between NOK and Akt, while constitutively active mutant of Akt apparently had a stronger interaction with NOK. NOK did not have an additive effect on insulin-mediated Akt activation. Overall, the results indicate that NOK might complex with Akt and directly activate PI3K/Akt signaling pathway.