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Previous Article | Next Article 
Identification of Proteins in the Postsynaptic Density Fraction
by Mass Spectrometry
Randall S.
Walikonis1,
Ole N.
Jensen2,
Matthias
Mann2,
D. William
Provance Jr3,
John A.
Mercer3, and
Mary B.
Kennedy1
1 Division of Biology, California Institute of
Technology, Pasadena, California 91125, 2 European
Molecular Biology Laboratory, D69012 Heidelberg, Germany, and
3 McLaughlin Research Institute, Great Falls, Montana
59405-4900
Our understanding of the organization of postsynaptic signaling
systems at excitatory synapses has been aided by the identification of
proteins in the postsynaptic density (PSD) fraction, a subcellular fraction enriched in structures with the morphology of PSDs. In this
study, we have completed the identification of most major proteins in
the PSD fraction with the use of an analytical method based on mass
spectrometry coupled with searching of the protein sequence databases.
At least one protein in each of 26 prominent protein bands from the PSD
fraction has now been identified. We found 7 proteins not previously
known to be constituents of the PSD fraction and 24 that had previously
been associated with the PSD by other methods. The newly identified
proteins include the heavy chain of myosin-Va (dilute myosin), a motor
protein thought to be involved in vesicle trafficking, and the
mammalian homolog of the yeast septin protein cdc10, which is important
for bud formation in yeast. Both myosin-Va and cdc10 are threefold to fivefold enriched in the PSD fraction over brain homogenates. Immunocytochemical localization of myosin-Va in cultured hippocampal neurons shows that it partially colocalizes with PSD-95 at synapses and
is also diffusely localized in cell bodies, dendrites, and axons. Cdc10
has a punctate distribution in cell bodies and dendrites, with some of
the puncta colocalizing with PSD-95. The results support a role for
myosin-Va in transport of materials into spines and for septins in the
formation or maintenance of spines.
Key words:
synaptic transmission; myosin; septins; vesicle
transport; signal transduction; multiprotein complex
Copyright © 2000 Society for Neuroscience 0270-6474/00/20114069-12$05.00/0
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