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  • Non-microtubule tubulin-based backbone and subordinate components of postsynaptic density lattices.

Non-microtubule tubulin-based backbone and subordinate components of postsynaptic density lattices.

Life science alliance (2021-05-20)
Tatsuo Suzuki, Nobuo Terada, Shigeki Higashiyama, Kiyokazu Kametani, Yoshinori Shirai, Mamoru Honda, Tsutomu Kai, Weidong Li, Katsuhiko Tabuchi
ABSTRACT

A purification protocol was developed to identify and analyze the component proteins of a postsynaptic density (PSD) lattice, a core structure of the PSD of excitatory synapses in the central nervous system. "Enriched"- and "lean"-type PSD lattices were purified by synaptic plasma membrane treatment to identify the protein components by comprehensive shotgun mass spectrometry and group them into minimum essential cytoskeleton (MEC) and non-MEC components. Tubulin was found to be a major component of the MEC, with non-microtubule tubulin widely distributed on the purified PSD lattice. The presence of tubulin in and around PSDs was verified by post-embedding immunogold labeling EM of cerebral cortex. Non-MEC proteins included various typical scaffold/adaptor PSD proteins and other class PSD proteins. Thus, this study provides a new PSD lattice model consisting of non-microtubule tubulin-based backbone and various non-MEC proteins. Our findings suggest that tubulin is a key component constructing the backbone and that the associated components are essential for the versatile functions of the PSD.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Protease Inhibitor Cocktail, for use with mammalian cell and tissue extracts, DMSO solution
Sigma-Aldrich
Anti-Internexin α Antibody, serum, Chemicon®
Sigma-Aldrich
Anti-Spectrin alpha chain (nonerythroid) Antibody, clone AA6, clone AA6, Chemicon®, from mouse
Sigma-Aldrich
Rabbit Anti-Goat IgG Antibody, Fc, HRP conjugate, 1.0 mg/mL, Chemicon®