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Aims The impact of long-term endurance sport participation (on top of a healthy lifestyle) on coronary atherosclerosis and acute cardiac events remains controversial. Methods and results The Master@Heart study is a well-balanced prospective observational cohort study. Overall, 191 lifelong master endurance athletes, 191 late-onset athletes (endurance sports initiation after 30 years of age), and 176 healthy non-athletes, all male with a low cardiovascular risk profile, were included. Peak oxygen uptake quantified fitness. The primary endpoint was the prevalence of coronary plaques (calcified, mixed, and non-calcified) on computed tomography coronary angiography. Analyses were corrected for multiple cardiovascular risk factors. The median age was 55 (50-60) years in all groups. Lifelong and late-onset athletes had higher peak oxygen uptake than non-athletes [159 (143-177) vs. 155 (138-169) vs. 122 (108-138) % predicted]. Lifelong endurance sports was associated with having >= 1 coronary plaque [odds ratio (OR) 1.86, 95% confidence interval (CI) 1.17-2.94], >= 1 proximal plaque (OR 1.96, 95% CI 1.24-3.11), >= 1 calcified plaques (OR 1.58, 95% CI 1.01-2.49), >= 1 calcified proximal plaque (OR 2.07, 95% CI 1.28-3.35), >= 1 non-calcified plaque (OR 1.95, 95% CI 1.12-3.40), >= 1 non-calcified proximal plaque (OR 2.80, 95% CI 1.39-5.65), and >= 1 mixed plaque (OR 1.78, 95% CI 1.06-2.99) as compared to a healthy non-athletic lifestyle. Conclusion Lifelong endurance sport participation is not associated with a more favourable coronary plaque composition compared to a healthy lifestyle. Lifelong endurance athletes had more coronary plaques, including more non-calcified plaques in proximal segments, than fit and healthy individuals with a similarly low cardiovascular risk profile. Longitudinal research is needed to reconcile these findings with the risk of cardiovascular events at the higher end of the endurance exercise spectrum. [GRAPHICS] . This study is supported by a research grant (T003717N) and R.W. is supported as a post-doctoral clinical researcher by the Fund for Scientific Research Flanders (FWO Vlaanderen), Brussels, Belgium The authors would like to thank the many staff members at the three sites for helping conduct this study. We would particularly like to thank the clinical research assistants Sofie van Soest, Dorien Vermeulen, and Daisy Thijs for their dedication and devoted efforts to include, test, and follow up participants. We would like to thank Ann Belmans for calculating the sample size and statistical advice. We would also like to thank Kris Van der Mieren (Sport- en keuringsartsen), Tom Teuglinckx (Sport- en keuringsartsen), Reynout Van Schuylenbergh (Triatlon Vlaanderen), Frank Glorieux (Cycling Vlaanderen), Paul Rowe (Sport Vlaanderen), Gert Van Goolen (Golazo), and Sven Nys (Golazo and godfather of Master@Heart) as members of the advisory committee for their insightful input on the design, the launch, and approach of the Master@Heart study.