Urbanisation alters biodiversity patterns and threatens to disrupt mutualistic interactions. Aside from pollination, however, little is known about how mutualisms change in cities. Our study aimed to assess how urbanisation affects the protective mutualism between ants and aphids, investigating potential behavioural changes in mutualistic ants and their implications for aphids in urban environments. To do so, we studied the protective mutualism between the pink tansy aphid (Metopeurum fuscoviride) and the black garden ant (Lasius niger) along an urbanisation gradient in Berlin, Germany. In nine locations along this gradient, we measured aphid colony dynamics and proxies for parasitism, quantified the investment of ants in tending aphids and conducted behavioural assays to test the aggressiveness of ant responses to a simulated attack on the aphids. We found that aphid colonies flourished and were equally tended by ants across the urbanisation gradient, with a consistent positive density dependence between aphid and ant numbers. However, ants from more urbanised sites responded more aggressively to the simulated attack. Our findings suggest that this protective mutualism is not only maintained in the city, but that ants might even rely more on it and defend it more aggressively, as other food resources may become scarce and more unpredictable with urbanisation. We thereby provide unique insights into this type of mutualism in the city, further diversifying the growing body of work on mutualisms across urbanisation gradients.