The purpose of the paper is to examine the mediating role of perceived organizational support (POS) in the relationship between work-related factors and job performance. A cross-sectional survey has been conducted among 323 healthcare professionals (medical officers and staff nurses) working in primary health centres of Tamilnadu, India to collect data using a questionnaire. Multiple regression analysis and Sobel test to analyse data. Results indicated that job autonomy, supervisor support and co-worker support partially influenced job performance through POS. The findings revealed the vital factors affecting the job performance of healthcare professionals specifically in a developing country like India. Based on the findings, the healthcare institutions need to enhance a supportive work environment to enhance the level of POS and consequently the job performance of healthcare professionals. © IAEME Publication.