The presynaptic particle web: ultrastructure, composition, dissolution, and reconstitution.
We report the purification of a presynaptic "particle web" consisting of approximately 50 nm pyramidally shaped particles interconnected by approximately 100 nm spaced fibrils. This is the "presynaptic grid" described in early EM studies. It is completely soluble above pH 8, but reconstitutes after dialysis against pH 6. Interestingly, reconstituted particles orient and bind PSDs asymmetrically. Mass spectrometry of purified web components reveals major proteins involved in the exocytosis of synaptic vesicles and in membrane retrieval. Our data support the idea that the CNS synaptic junction is organized by transmembrane adhesion molecules interlinked in the synaptic cleft, connected via their intracytoplasmic domains to the presynaptic web on one side and to the postsynaptic density on the other. The CNS synaptic junction may therefore be conceptualized as a complicated macromolecular scaffold that isostatically bridges two closely aligned plasma membranes.
Phillips
G R
GR
The Corinne Goldsmith Dickinson Center for Multiple Sclerosis, Department of Neurology and The Fishberg Research Center for Neurobiology, New York, NY 10029, USA.
Huang
J K
JK
Wang
Y
Y
Tanaka
H
H
Shapiro
L
L
Zhang
W
W
Shan
W S
WS
Arndt
K
K
Frank
M
M
Gordon
R E
RE
Gawinowicz
M A
MA
Zhao
Y
Y
Colman
D R
DR
D000818:Animals; D000906:Antibodies; D015820:Cadherins / Q000032:analysis / Q000276:immunology; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D002966:Clathrin / Q000032:analysis / Q000276:immunology; D033922:Clathrin Heavy Chains; D034281:Dynamins; D020558:GTP Phosphohydrolases / Q000032:analysis / Q000276:immunology; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis / Q000276:immunology; D008297:Male; D008565:Membrane Proteins / Q000032:analysis / Q000276:immunology; D016253:Microscopy, Immunoelectron; D051938:Munc18 Proteins; D018995:Myosin Heavy Chains / Q000032:analysis / Q000276:immunology; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000276:immunology; D016900:Neurofilament Proteins / Q000032:analysis / Q000276:immunology; D017729:Presynaptic Terminals / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D050765:Qa-SNARE Proteins; D011817:Rabbits; D051381:Rats; D013049:Spectrin / Q000032:analysis / Q000276:immunology; D016704:Synapsins / Q000032:analysis / Q000276:immunology; D013572:Synaptic Vesicles / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D050825:Synaptosomal-Associated Protein 25; D033921:Vesicular Transport Proteins
D000906:Antibodies; D015820:Cadherins; D002966:Clathrin; D018840:HSP70 Heat-Shock Proteins; D008565:Membrane Proteins; D051938:Munc18 Proteins; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D050765:Qa-SNARE Proteins; C494210:Snap25 protein, rat; D016704:Synapsins; D050825:Synaptosomal-Associated Protein 25; D033921:Vesicular Transport Proteins; D033922:Clathrin Heavy Chains; D013049:Spectrin; D020558:GTP Phosphohydrolases; D018995:Myosin Heavy Chains; D034281:Dynamins