In vitro motility of AtKCBP, a calmodulin-binding kinesin protein of Arabidopsis

AtKCBP is a calcium-dependent calmodulin-binding protein from Arabidopsis that contains a conserved kinesin microtubule motor domain, Calmodulin has been shown previously to bind to heavy chains of the unconventional myosins, where it is required for in vitro motility of brush border myosin 1, but AtKCBP is the first kinesin-related heavy chain reported to be capable of binding specifically to calmodulin. Other kinesin proteins have been identified in Arabidopsis, but none of these binds to calmodulin, and none has been demonstrated to be a microtubule motor. We have tested bacterially expressed AtKCBP for the ability to bind microtubules to a glass surface and induce gliding of microtubules across the glass surface. We find that AtKCBP is a microtubule motor protein that moves on microtubules toward the minus ends, with the opposite polarity as kinesin, In the presence of calcium and calmodulin, AtKCBP no longer binds microtubules to the coverslip surface, This contrasts strikingly with the requirement of calmodulin for in vitro motility of brush border myosin I. Calmodulin could regulate AtKCBP binding to microtubules in the cell by inhibiting the binding of the motor to microtubules. The ability to bind to calmodulin provides an evolutionary link between the kinesin and myosin motor proteins, but our results indicate that the mechanisms of interaction and regulation of kinesin and myosin heavy chains by calmodulin are likely to differ significantly.