Characterization of microtubule binding domains in the Arabidopsis kinesin-like calmodulin binding protein.
The kinesin-like calmodulin binding protein (KCBP) is a new member of the kinesin superfamily that appears to be present only in plants. The KCBP is unique in its ability to interact with calmodulin in a Ca2+-dependent manner. To study the interaction of the KCBP with microtubules, we expressed different regions of the Arabidopsis KCBP and used the purified proteins in cosedimentation assays with microtubules. The motor domain with or without the calmodulin binding domain bound to microtubules. The binding of the motor domain containing the calmodulin binding region to microtubules was inhibited by Ca2+-calmodulin. This Ca2+-calmodulin regulation of motor domain interactions with microtubules was abolished in the presence of antibodies specific to the calmodulin binding region. In addition, the binding of the motor domain lacking the calmodulin binding region to microtubules was not inhibited in the presence of Ca2+-calmodulin, suggesting an essential role for the calmodulin binding region in Ca2+-calmodulin modulation. Results of the cosedimentation assays with the N-terminal tail suggest the presence of a second microtubule binding site on the KCBP. However, the interaction of the N-terminal tail region of the KCBP with microtubules was insensitive to ATP. These data on the interaction of the KCBP with microtubules provide new insights into the functioning of the KCBP in plants.
Narasimhulu
S B
SB
Department of Biology and Program in Cell and Molecular Biology, Colorado State University, Fort Collins, Colorado 80523, USA.
Reddy
A S
AS
D000818:Animals; D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins; D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002147:Calmodulin / Q000494:pharmacology; D002148:Calmodulin-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D002417:Cattle; D016547:Kinesins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D008870:Microtubules / Q000187:drug effects / Q000502:physiology*; D010940:Plant Proteins / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D014404:Tubulin / Q000302:isolation & purification / Q000378:metabolism
D029681:Arabidopsis Proteins; D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; C099551:KCBP protein, Arabidopsis; D010940:Plant Proteins; D011994:Recombinant Proteins; D014404:Tubulin; D016547:Kinesins; D002118:Calcium