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Calcium/calmodulin-dependent inhibition of microtubule assembly by brain synaptic junction

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Abstract

The effect of synaptic junction (SJ) on microtubule assembly was examined. After preincubation with ATP at 37°C, rat SJ decreased the initial velocity and the extent of the porcine brain microtubule assembly (initiated by the addition of GTP) in a Ca2+/calmodulin (CaM)-dependent manner. The degree of the inhibition reached 35% of the control assembly (0-min preincubation) after 20-min preincubation with ATP. There was no inhibition either with heat-treated SJ, at 0°C, or in the presence of EGTA or W-7 (CaM antagonist). The inhibition was due neither to protease(s) nor CaM contaminating the preparations. Free Ca2+ concentration level required for the inhibition of microtubule assembly was 10−6 M. Phosphorylation of microtubule proteins was inhibited by SJ in a Ca2+/CaM-dependent manner, and the inhibition occurred in a physiological increase range of intracellular Ca2+ concentration (10−6M) The heat-treated SJ caused no inhibition. The result suggested that the microtubule assembly in the postsynaptic region was regulated by a Ca2+/CaM-dependent protein kinase associated with SJ; i. e., major postsynaptic density protein.

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Abbreviations

CaM:

calmodulin

DTT:

dithiothreitol

MAPs:

microtubule-associated proteins

MES:

2-(N-morphorino)ethanesulfonic acid

mPSDp:

major postsynaptic density protein

PSD:

postsynaptic density

SDS PAGE:

sodium dodecyl sulfate polyacrylamide gel electrophoresis

W-7:

N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride

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Authors and Affiliations

  1. Department of Biochemistry, Nagoya City University Medical School, Mizuho-ku, 467, Nagoya, Japan

    Tatsuo Suzuki & Ryo Tanaka

  2. Department of Functional Polymer Science Faculty of Textile Science and Technology, Shinshu University, 386, Ueda, Japan

    Toshihiro Fujii

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  1. Tatsuo Suzuki
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  2. Toshihiro Fujii
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Suzuki, T., Fujii, T. & Tanaka, R. Calcium/calmodulin-dependent inhibition of microtubule assembly by brain synaptic junction. Neurochem Res 11, 543–555 (1986). https://doi.org/10.1007/BF00965324

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