Heavy chain of cytoplasmic dynein is a major component of the postsynaptic density fraction.
A protein with an apparent molecular size of 490 kDa was found in the postsynaptic density (PSD) fraction isolated from porcine cerebral cortices and rat forebrains, and this 490 kDa protein accounted for approximately 3% of the total protein of these samples. Matrix-assisted laser desorption ionization-time of flight mass spectrometric and Western blotting analyses consistently indicated that this 490 kDa protein consisted primarily of the heavy chain of cytoplasmic dynein (cDHC). Immunocytochemical analyses showed that cDHC was found in 92% and 89% of the phalloidin-positive protrusions that were themselves associated with discrete clusters of synaptophysin, a presynaptic terminal marker, and PSD-95, a postsynaptic marker, on neuronal processes, respectively. Quantitative Western blotting analyses of various subcellular fractions isolated from porcine cerebral cortices and rat forebrains further showed that not only the heavy but also the intermediate chains of dynein are enriched in the PSD fraction. Cytoplasmic dynein is a microtubule-associated motor protein complex that drives the movement of various cargos toward the minus ends of microtubules and plays many other diverse functions in the cell. Our results that cDHC is a major component of the PSD fraction, that both dynein heavy and intermediate chains are enriched in the PSD fraction and that cDHC is present in dendritic spines raise the possibilities that cytoplasmic dynein may play structural and functional roles in the postsynaptic terminal.
Cheng
Huei-Hsuan
HH
Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan, Republic of China.
Liu
Szu-Heng
SH
Lee
Hui-Cheng
HC
Lin
Ya-Shiuan
YS
Huang
Zu-Han
ZH
Hsu
Cheng-I
CI
Chen
Yu-Chie
YC
Chang
Yen-Chung
YC
D000818:Animals; D015153:Blotting, Western; D001923:Brain Chemistry; D003593:Cytoplasm / Q000737:chemistry*; D004398:Dyneins / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D007150:Immunohistochemistry; D009419:Nerve Tissue Proteins / Q000737:chemistry; D009474:Neurons / Q000737:chemistry; D050765:Qa-SNARE Proteins / Q000032:analysis; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013347:Subcellular Fractions / Q000737:chemistry*; D013552:Swine; D016708:Synaptophysin / Q000032:analysis
D009419:Nerve Tissue Proteins; D050765:Qa-SNARE Proteins; D016708:Synaptophysin; C033652:postsynaptic density proteins; D004398:Dyneins