The Enlightenment has long been defined as an age of expanding knowledge. Practices of collection, classification and display of objects, which intensified and spread along with the global extension of European empires and commercial networks, meant that Enlightenment intellectual aspiration became global in scope. This article focuses on the colonial collections of the Professor of Natural History at the University of Edinburgh, the Rev. Dr John Walker, who was also the keeper of the university’s natural history museum. This article studies in particular the actors involved in the movement of a large collection of objects from the Hudson’s Bay Company. The collection was provided by an employee of the Company, Andrew Graham who also penned a manuscript about the artefacts and the people inhabiting Rupert’s Land. Graham’s collecting network included other traders, First Nation and Inuit actors and European-based naturalists. The article highlights the importance of conferring historical agency on a diverse cast of figures in the mobile formation and communication of colonial knowledge about humanity. It argues, however, that this movement of knowledge was not frictionless but was conditioned by uneven power relations and violence.