Skip to Content
Merck
  • Compounds activating VCP D1 ATPase enhance both autophagic and proteasomal neurotoxic protein clearance.

Compounds activating VCP D1 ATPase enhance both autophagic and proteasomal neurotoxic protein clearance.

Nature communications (2022-07-17)
Lidia Wrobel, Sandra M Hill, Alvin Djajadikerta, Marian Fernandez-Estevez, Cansu Karabiyik, Avraham Ashkenazi, Victoria J Barratt, Eleanna Stamatakou, Anders Gunnarsson, Timothy Rasmusson, Eric W Miele, Nigel Beaton, Roland Bruderer, Yuehan Feng, Lukas Reiter, M Paola Castaldi, Rebecca Jarvis, Keith Tan, Roland W Bürli, David C Rubinsztein
ABSTRACT

Enhancing the removal of aggregate-prone toxic proteins is a rational therapeutic strategy for a number of neurodegenerative diseases, especially Huntington's disease and various spinocerebellar ataxias. Ideally, such approaches should preferentially clear the mutant/misfolded species, while having minimal impact on the stability of wild-type/normally-folded proteins. Furthermore, activation of both ubiquitin-proteasome and autophagy-lysosome routes may be advantageous, as this would allow effective clearance of both monomeric and oligomeric species, the latter which are inaccessible to the proteasome. Here we find that compounds that activate the D1 ATPase activity of VCP/p97 fulfill these requirements. Such effects are seen with small molecule VCP activators like SMER28, which activate autophagosome biogenesis by enhancing interactions of PI3K complex components to increase PI(3)P production, and also accelerate VCP-dependent proteasomal clearance of such substrates. Thus, this mode of VCP activation may be a very attractive target for many neurodegenerative diseases.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Cycloheximide, from microbial, ≥94% (TLC)
Sigma-Aldrich
Anti-Huntingtin Protein Antibody, a.a. 181-810, clone 1HU-4C8, ascites fluid, clone 1HU-4C8, Chemicon®
PreScission Protease, Cytiva 27-0843-01
Sigma-Aldrich
Anti-Ubiquitinylated proteins Antibody, clone FK2, clone FK2, Upstate®, from mouse
Sigma-Aldrich
Anti-Spinocerebellar Ataxia Type 3 Antibody, clone 1H9, ascites fluid, clone 1H9, Chemicon®
Sigma-Aldrich
Anti-Polyglutamine-Expansion Diseases Marker Antibody, clone 5TF1-1C2, ascites fluid, clone 5TF1-1C2, Chemicon®
Sigma-Aldrich
Anti-Puromycin Antibody, clone 12D10, clone 12D10, from mouse
Sigma-Aldrich
Anti-Actin antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Z-Leu-Leu-Leu-al, ≥90% (HPLC)
Sigma-Aldrich
ANTI-FLAG® M2 antibody, Mouse monoclonal, 1.0-1.2 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)