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The Arabidopsis calcineurin B‐like calcium sensor proteins (AtCBLs) interact with a group of serine‐threonine protein kinases (AtCIPKs) in a calcium‐dependent manner. Here we identify a 24 amino acid domain (NAF domain) unique to these kinases as being required and sufficient for interaction with all known AtCBLs. Mutation of conserved residues either abolished or significantly diminished the affinity of AtCIPK1 for AtCBL2. Comprehensive two‐hybrid screens with various AtCBLs identified 15 CIPKs as potential targets of CBL proteins. Database analyses revealed additional kinases from Arabidopsis and other plant species harbouring the NAF interaction module. Several of these kinases have been implicated in various signalling pathways mediating responses to stress, hormones and environmental cues. Full‐length CIPKs show preferential interaction with distinct CBLs in yeast and in vitro assays. Our …