A key success factor for a successful company in a dynamic environment is effective and efficient information technology (IT) supporting business strategies and processes. Organizations that successfully align their business strategy and their IT strategy outperform their non-aligned peers (Chan et al., 1997). In recent surveys IT executives consistently name IT to Business alignment their top-concern. The alignment between business needs and IT capabilities is therefore still a prominent area of concern. This paper aims to contribute to the understanding of the alignment challenge by exploring the relationship between organizational culture and the maturity of business and IT alignment (BIA). The paper relies on the X-model of organizational culture (Smit et al., 2008) and Luftman’s framework for measuring BIA maturity (Luftman, 2000) to explore the relationships between these concepts A quantitative study was conducted in a middle-sized logistics service provider using a questionnaire that was derived from the afore-mentioned models. The results support the notion that there is a relationship between organizational culture and BIA maturity, especially on the variables ‘governance’, ‘partnership’ and ‘skills’. Further research is required to determine causality in these relationships.