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Identification of proteins in the postsynaptic density fraction by mass spectrometry.
- Source :
-
The Journal of neuroscience : the official journal of the Society for Neuroscience [J Neurosci] 2000 Jun 01; Vol. 20 (11), pp. 4069-80. - Publication Year :
- 2000
-
Abstract
- 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.
- Subjects :
- Amino Acid Sequence
Animals
Cell Cycle Proteins analysis
Cell Cycle Proteins biosynthesis
Cell Cycle Proteins genetics
Cells, Cultured
Cloning, Molecular
DNA, Complementary biosynthesis
DNA, Complementary chemistry
Dendrites chemistry
Hippocampus chemistry
Hippocampus cytology
Hydrolysis
Immunoblotting
Immunohistochemistry
Intermediate Filament Proteins analysis
Mass Spectrometry
Molecular Sequence Data
Neurons chemistry
Peptides analysis
Rats
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Trypsin
Myosin Heavy Chains
Myosin Type V
Nerve Tissue Proteins analysis
Synapses chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1529-2401
- Volume :
- 20
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- The Journal of neuroscience : the official journal of the Society for Neuroscience
- Publication Type :
- Academic Journal
- Accession number :
- 10818142