Mycobacterium tuberculosis (Mtb) is the causative agent of tuberculosis (TB), capable of manipulating and circumventing the host's immune system to establish infection. Ubiquitination plays a crucial role in the host's response to pathogens; however, the global alterations in protein ubiquitination during Mtb infection remain poorly understood. To elucidate the regulatory roles of ubiquitination in the immune response to Mtb, we investigated the ubiquitome of human macrophages following Mtb infection.