Kinases and pseudokinases play diverse roles in cellular signalling, and dysfunction of these pathways is linked to numerous human diseases. We recently identified patients with histories of inflammatory disease, who each had mutations in either the PSKH1 kinase, or non-catalytic PSKH2 pseudokinase. These human proteins are poorly characterised and have been termed ‘dark’ kinases by the NIH. Our interest was piqued by our identification of novel interactors of PSKH1/2 that are integral to TNF signalling using mass spectrometry. TNF signalling pathways regulate the immune system, but are also widely implicated in inflammatory pathology, asserting the disease relevance of these ‘dark’ kinases. PSKH1/2 are members of the Ca2+/Calmodulin-dependent protein kinase family. Excitingly, we have discovered a new mode of Ca2+/Calmodulin activation within the kinase domain, that is distinct from those previously reported. Adding to the intrigue, the patient mutations we have identified are located near the putative Calmodulin binding site, which we predict could impact the activation mechanism. Together, our discoveries present a unique opportunity to unravel how these ‘dark’ kinases are activated and regulated in cells. We also establish PSKH1/2 as exciting new candidates for the development of anti-TNFs to treat the often life-threating inflammatory disease.