Objectives: The purpose of this study was to examine measures of cardiac and respiratory activity when participants at different levels of skill development performed a golf putting task under different attentional focus instructions. Putting performance and self-reports of attentional focus were also examined. Methods: Novice (n = 18), experienced (n = 16), and elite golfers (n = 16) attempted 2.4 m straight putts under a baseline (no instruction) condition and when instructed to focus attention on a process goal, a performance goal, an outcome goal, or to trust the body to perform the skill. Results: Compared to novice golfers, the experienced and elite golfers showed better performance and reduced heart rate (HR), greater heart rate variability (HRV), pronounced HR deceleration prior to the putt, and a greater tendency to exhale prior to the putt. The attentional focus instructions also influenced HR and putting performance. Conclusions: The results show that athletes at different skill levels differ in their performance and focus of attention while performing a motor task.