Assessment of palmitoylation status and cycles on PSD 95 in neurons and in vivo

• Autorzy: Fukata Y. , Yokoi N. , Kobayashi K. , Fukata M.
• Języki publikacji: EN

Abstrakty

EN

INTRODUCTION: Precise synaptic function requires spatio-temporally regulated protein localization. Protein palmitoylation, a reversible lipid modification, represents one such mechanism. Although numerous synaptic palmitoylated proteins have been identified, the physiological importance of their palmitoylation remains incompletely understood due to the lack of quantitative information. AIM(S): To determine the actual palmitoylation stoichiometry and state (for example, mono-, di-, tri-) of representative synaptic proteins in the rat brain, and to examine how dynamically the palmitoyl-turnover on proteins is regulated. METHOD(S): We used recently developed acyl-PEGyl exchange gel‑shift (APEGS) assay to profile palmitoylation stoichiometry of synaptic proteins and their dynamic changes, especially for PSD-95, in rat cultured hippocampal neurons and in rat brain. RESULTS: We found that individual palmitoylated proteins have the distinct palmitoylation site occupancy and the kinetics in rat cultured hippocampal neurons. Unexpectedly, palmitate on synaptic proteins did not all turn over. Of particular importance however is uniquely robust and dynamic palmitoylation for a postsynaptic scaffold PSD-95. In young neurons the stoichiometry of PSD‑95 palmitoylation was about 60% with the rapid palmitate cycling, whereas palmitate cycling on PSD-95 significantly decelerated accompanied by the increased stoichiometry in neurons and in vivo. Furthermore, we found that the sensitivity against recently discovered PSD-95 depalmitoylating enzyme, ABHD17, well correlated with the speed of palmitoyl cycling and cluster formation of PSD-95. CONCLUSIONS: This study suggests that the palmitoylation stoichiometry and kinetics of PSD-95 could be tightly controlled in response to the physiological contexts during synapse development by the specific palmitoylating/depalmitoylating enzymes. FINANCIAL SUPPORT: The Ministry of Education, Culture, Sports, Science and Technology – Japan (15H04279, 15H01299)

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2017
• Tom: 77
• Numer: Suppl.1
• Opis fizyczny: p.107

Twórcy

autor

Fukata Y.

• Division of Membrane Physiology, Department of Molecular and Cellular Physiology, Okazaki, Japan
• Department of Physiological Sciences, School of Life Science, SOKENDAI, Okazaki, Japan

autor

Yokoi N.

• Division of Membrane Physiology, Department of Molecular and Cellular Physiology, Okazaki, Japan
• Department of Physiological Sciences, School of Life Science, SOKENDAI, Okazaki, Japan

autor

Kobayashi K.

• Department of Physiological Sciences, School of Life Science, SOKENDAI, Okazaki, Japan
• Section of Viral Vector Development, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki, Japan

autor

Fukata M.

• Division of Membrane Physiology, Department of Molecular and Cellular Physiology, Okazaki, Japan
• Department of Physiological Sciences, School of Life Science, SOKENDAI, Okazaki, Japan