Alienation is an important topic in organizational sciences of the west. Employee alienation is emerging rapidly in china’s workplace. A large number of studies have linked employee alienation to both individual differences (e.g., achievement, work ethic) and structural predictors such as role stressors, leader dimensions (e.g., supportive leadership), job design (e.g., task variety), and work context (e.g., formalization, centralization). However the alienation’s nomological network is still somewhat unclear. In this study, a negative organizational factor—perceived organization’s political climate and a new psychological mediation—self-determination motivation of work are considered to explore the forming mechanism of work alienation in China’s context. In the present study, the self-report data were collected from 226 employee participants of 8 enterprises and institutions in Shanghai, China, by convenient sampling survey using Perceived General Organization’s Political Behavior Scale, Work Extrinsic and Intrinsic Motivation Scale and Work Alienation Scale. Correlation analysis and the Structural Equation Modeling (SEM) were used to reveal the relationships of the variables. The Work Alienation Scale contains two dimensions: personal alienation and social alienation. The results showed that: (1) organization’s political climate perceived by Chinese employees was positively correlated with personal alienation (r = .685, p < .01) and social alienation (r = .628, p < .01). And it was negatively correlated with self-determination motivation of work (r = -.322, p < .01). (2) Self-determination motivation of work was negatively correlated with personal alienation (r = -.413, p < .01) and social alienation (r = -.278, p < .01). (3) The path coefficients of SEM were significant from perceived organization’s political climate to personal alienation (.65, p < .001), social alienation (.65, p < .001) and self-determination motivation of work (-.33, p < .001). (4) The path coefficient of SEM was significant from self-determination motivation of work to personal alienation (-.21, p < .001), while to social alienation was weak and not significant (-.08, p > .05). SEM showed that the self-determination motivation of work played a partially mediating role in the relationship between perceived organization’s political climate and employee’s personal alienation. The present study not only examined the relationships between perceived organization’s political climate and work alienation, but also found the mediating role of self-determination motivation of work, which expanded the related areas and enriched the original theory. As a negative interactive organizational factor, the organization’s political climate is an important predictor causing feelings of work alienation. Workplace full of political behaviors can cause employees negative senses of separation or estrangement extending to one’s self-image (personal alienation) and social relationships (social alienation). And the self-determination motivation of work is a psychological mediation mechanism which understands the formation of employee’s personal alienation to work activities. That is to say, the employee’s personal alienation experience to work is partly due to the weakening effect of the organizational political atmosphere on employee’s self-determination motivation of work. But the employee’s social alienation experience to social relationship is mainly due to the direct effect of the organizational political atmosphere perceived by employees in the working world.