Purpose: Smoking and physical inactivity are two major health problems that concern the public. Although the relationship between physical activity level and smoking status is defined in different study groups, this relationship is not clearly defined in health care professionals who are thought to be a guide for community health. Therefore, our study aimed to examine the relationship between smoking and physical activity level in healthcare professionals. Methods: A total of 193 healthcare professionals (median age; 38 [interquartile range-IQR; 34-45] years, body mass index; 24 [IQR; 21-27] kg/m2), 102 smokers and 91 non-smokers, who worked in an educational research hospital, participated in the study. Physical activity was assessed using the International Physical Activity Questionnaire. Smoking history was taken in the smoker group, and the Fagerstrom Test for Nicotine Dependence was administered to determine the nicotine addiction level. Results: The demographics of the groups were similar (p>0.05). Walking activity, moderate physical activity and total physical activity scores were significantly lower in smokers than non-smokers (p<0.001, p=0.021, and p=0.001, respectively). The total physical activity score in the smoking group was showed a moderate negative correlation with the nicotine dependence and the amount of cigarette consumption (r=-0.462 and r=-0.483, respectively, p<0.05). Conclusion: The level of physical activity in smokers' health professionals was found to be low. As the amount of cigarette consumption and nicotine dependence increases, the level of physical activity decreases. In this crucial population, further studies should be undertaken to improve both smoking cessation and physical activity.