Background. The link between organizational culture, organizational trust, job satisfaction, and team-based practice among nurse practitioners (NPs) has not been examined simultaneously. Aim. To identify the effects of organizational culture, organizational trust, and other factors on NPs such as job satisfaction and team-based practice. Methods. We used a cross-sectional design with a national sample. Data were collected using an online survey of 1,100 NPs working in acute care settings. The survey included demographic and working characteristics, the Organizational Culture Scale, the Organizational Trust Scale, the Misener Nurse Practitioner Job Satisfaction Scale (MNPJSS), and the NP-physician relations (NP-PR) subscale of the Nurse Practitioner Primary Care Organizational Climate (NP-PCOCQ). Multiple regression analysis with a stepwise selection method explored potential factors that influence job satisfaction and team-based practice. Results. A learning environment, psychological safety, senior leadership support, commitment to the organization, and the organizational culture and trust were positively associated with higher job satisfaction, which accounted for 49.2% of the variance in NPs’ job satisfaction. Organizational trust, commitment to the organization, and learning environment promoted better team-based practice significantly. Also, NPs working a fixed shift pattern showed higher levels of team-based practice. These factors accounted for 23.66% of variances in team-based practice. Conclusion. Organizational culture and organizational trust affect the job satisfaction and team-based practice of NPs in acute care practices. Implications for Nursing Management. Acute care hospitals are encouraged to develop policies to enhance a learning environment, a supportive organizational culture, and trust in NPs’ practice.