Characterization of granular particles isolated from postsynaptic densities.
We describe here the isolation and biochemical characterization of a population of protein aggregates from the postsynaptic density (PSD) prepared from pig cerebral cortex. The protein constituents of these aggregates are linked together primarily by disulfide bonds. Negative staining electron microscopy revealed that the isolated protein aggregates were granular objects with an average outside diameter of approximately 21 nm and with small protrusions on their surface. The major constituents of the isolated granular aggregates consist of tubulin and an unidentified protein of 70 kDa in size. Small amounts of the alpha subunit of calcium/calmodulin-dependent protein kinase II and subunits of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and NMDA subtypes of glutamate receptors were also detected by immunoblotting. Actin, however, was not found in these granular aggregates. We propose that these granular protein aggregates correspond to the approximately 20-nm-diameter granular particles of the PSD on the basis of their biochemical and morphological characteristics. The spatial arrangement of these granular aggregates relative to other components of the postsynaptic terminal is also postulated here.
Lai
S L
SL
Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan, Republic of China.
Ling
S C
SC
Kuo
L H
LH
Shu
Y C
YC
Chow
W Y
WY
Chang
Y C
YC
D000818:Animals; D002540:Cerebral Cortex / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D046911:Macromolecular Substances; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000648:ultrastructure; D017470:Receptors, Glutamate / Q000737:chemistry; D013347:Subcellular Fractions / Q000737:chemistry / Q000648:ultrastructure; D013552:Swine; D013569:Synapses / Q000737:chemistry*
D004220:Disulfides; D046911:Macromolecular Substances; D009419:Nerve Tissue Proteins; D017470:Receptors, Glutamate