Characterization of the microtubule-binding activity of kinesin-like calmodulin binding protein from Dunaliella salina.

Although the C-terminal motor and the N-terminal myosin-like domains of KCBP in Dunaliella salina (DsKCBP) are implicated in interaction with the microtubules, its microtubule binding property has not been addressed. It has been shown that several calmodulin isoforms suppress the microtubule binding activity of KCBP, but whether the calmodulin-like protein (CLP) has this ability remains unknown. The results of our previous study showed that there are two microtubule binding sites in DsKCBP, motor domain at the C-terminus and MyTH4-FREM at the N-terminus. In the present study, MyTH4, without the companion of FERM, was identified as the minimal domain responsible for interaction with the microtubules in the N-terminal of DsKCBP. CLP interacted with the calmodulin-binding domain of DsKCBP in the presence of Ca(2+), and inhibited the microtubule-binding activity of motor domain but not MyTH4 domain. Furthermore, MyTH4 domain in the N-terminus of DsKCBP was responsible for binding to the microtubules, and had 10-fold weaker affinity to the microtubules than the motor domain.