The tribological behavior of dry lubricants depends on their mechanical and physicochemical environment, making it difficult to predict in practice. Discrete Element Method-based modeling has been one successful approach to provide valuable insight into the tribology of dry lubricated contacts. However, it requires well-defined interactions between discrete elements, in particular between those simulating different materials. Measuring the properties governing those interactions, such as the work of adhesion (W), is therefore critical. The present work describes a method for measuring the W between AISI440C steel and MoS 2-based coatings used in spacecraft. Using Atomic Force Microscopy local asperity and adhesion measurements, the W between steel microbeads and MoS 2 coatings is determined at different stages in its wear life. The distributions of W values in the worn coatings and pristine coatings agree well with earlier Time-of-Flight Secondary Ion Mass Spectroscopy studies on the physicochemistry of the samples, as well as contact angle measurements. Additional measurements between the same materials on a ball bearing from a real life-test unit of a spacecraft instrument also show a similar W distribution, suggesting that the approach used here provides relevant data for use in numerical simulations.