This paper contributes to the calling literature by using self-determination theory – a theory that makes distinctions between different types of motivation –in order to gain a better understanding of how enacting a calling may relate both positively and negatively to well-being. We use a daily diary method novel to the calling field and a sample with a distinctive calling, Church of England clergy. We expect daily calling enactment to relate positively to daily well-being via more autonomous forms of motivation (intrinsic and identified motivation) and negatively via less autonomous forms (introjected motivation). Furthermore, we consider how the relationship between calling enactment and motivation may be moderated by perceived competence. The hypotheses were tested using multi-level structural equation modelling. There was strong support for calling enactment relating positively to well-being, and this relationship was fully mediated by intrinsic and identified motivation; the hypothesized negative pathway, from calling enactment, to introjected motivation, to well-being, was not supported. However, perceived competence was found to moderate some of the relationships between calling and the motivation types, whereby at high levels of competence calling enactment is linked to lower introjected motivation.