Engineering of a novel Ca2+-regulated kinesin molecular motor using a calmodulin dimer linker

► Engineered kinesin-M13 and calmodulin involving single cysteine were prepared. ► CaM mutant was cross-linked to dimer by bifunctional thiol reactive reagent. ► Kinesin-M13 was dimerized via CaM dimer in the presence of calcium. ► Function of the engineered kinesin was regulated by a Ca2+-calmodulin dimer linker.