Sökning: onr:"swepub:oai:DiVA.org:liu-184797" >
Comprehensive Targe...
Comprehensive Target Screening and Cellular Profiling of the Cancer-Active Compound b-AP15 Indicate Abrogation of Protein Homeostasis and Organelle Dysfunction as the Primary Mechanism of Action
-
- Gubat, Johannes, 1986- (författare)
- Linköpings universitet,Avdelningen för klinisk kemi och farmakologi,Medicinska fakulteten
-
- Selvaraju, Karthik, 1983- (författare)
- Linköpings universitet,Avdelningen för klinisk kemi och farmakologi,Medicinska fakulteten
-
- Sjöstrand, Linda (författare)
- Linköpings universitet,Avdelningen för klinisk kemi och farmakologi,Medicinska fakulteten
-
visa fler...
-
- Kumar Singh, Dhananjay (författare)
- Linköpings universitet,Institutionen för biomedicinska och kliniska vetenskaper,Medicinska fakulteten,Department of Pharmacy, Central University of South Bihar, Gaya, India
-
- Turkina, Maria V, 1973- (författare)
- Linköpings universitet,Avdelningen för cellbiologi,Medicinska fakulteten
-
- Schmierer, Bernhard (författare)
- Karolinska Institutet,Department of Medical Biochemistry and Biophysics, Division of Chemical Biology, Karolinska Institutet, Stockholm, Sweden
-
- Sabatier, Pierre (författare)
- Department of Medical Biochemistry and Biophysics, Division of Physiological Chemistry I, Karolinska Institutet, Stockholm, Sweden
-
- Zubarev, Roman A. (författare)
- Karolinska Institutet,Department of Medical Biochemistry and Biophysics, Division of Physiological Chemistry I, Karolinska Institutet, Stockholm, Sweden; Department of Pharmacological and Technological Chemistry, I.M. Sechenov First Moscow State Medical University, Moscow, Russia
-
- Linder, Stig, 1954- (författare)
- Linköpings universitet,Avdelningen för klinisk kemi och farmakologi,Medicinska fakulteten,Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden
-
- D´arcy, Pádraig, 1978- (författare)
- Karolinska Institutet,Linköpings universitet,Avdelningen för klinisk kemi och farmakologi,Medicinska fakulteten
-
visa färre...
-
(creator_code:org_t)
- 2022-04-22
- 2022
- Engelska.
-
Ingår i: Frontiers in Oncology. - : Frontiers Media SA. - 2234-943X. ; 12
- Relaterad länk:
-
https://doi.org/10.3...
-
visa fler...
-
https://liu.diva-por... (primary) (Raw object)
-
https://urn.kb.se/re...
-
https://doi.org/10.3...
-
http://kipublication...
-
visa färre...
Abstract
Ämnesord
Stäng
- Dienone compounds have been demonstrated to display tumor-selective anti-cancer activity independently of the mutational status of TP53. Previous studies have shown that cell death elicited by this class of compounds is associated with inhibition of the ubiquitin-proteasome system (UPS). Here we extend previous findings by showing that the dienone compound b-AP15 inhibits proteasomal degradation of long-lived proteins. We show that exposure to b-AP15 results in increased association of the chaperones VCP/p97/Cdc48 and BAG6 with proteasomes. Comparisons between the gene expression profile generated by b-AP15 to those elicited by siRNA showed that knock-down of the proteasome-associated deubiquitinase (DUB) USP14 is the closest related to drug response. USP14 is a validated target for b-AP15 and we show that b-AP15 binds covalently to two cysteines, Cys203 and Cys257, in the ubiquitin-binding pocket of the enzyme. Consistent with this, deletion of USP14 resulted in decreased sensitivity to b-AP15. Targeting of USP14 was, however, found to not fully account for the observed proteasome inhibition. In search for additional targets, we utilized genome-wide CRISPR/Cas9 library screening and Proteome Integral Solubility Alteration (PISA) to identify mechanistically essential genes and b-AP15 interacting proteins respectively. Deletion of genes encoding mitochondrial proteins decreased the sensitivity to b-AP15, suggesting that mitochondrial dysfunction is coupled to cell death induced by b-AP15. Enzymes known to be involved in Phase II detoxification such as aldo-ketoreductases and glutathione-S-transferases were identified as b-AP15-targets using PISA. The finding that different exploratory approaches yielded different results may be explained in terms of a “target” not necessarily connected to the “mechanism of action” thus highlighting the importance of a holistic approach in the identification of drug targets. We conclude that b-AP15, and likely also other dienone compounds of the same class, affect protein degradation and proteasome function at more than one level.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin -- Cancer och onkologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Clinical Medicine -- Cancer and Oncology (hsv//eng)
Nyckelord
- b-AP15
- proteasome inhibitor
- mitochondrial dysfunction
- dienone
- Michael acceptor
- target screening
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
Hitta via bibliotek
Till lärosätets databas
- Av författaren/redakt...
-
Gubat, Johannes, ...
-
Selvaraju, Karth ...
-
Sjöstrand, Linda
-
Kumar Singh, Dha ...
-
Turkina, Maria V ...
-
Schmierer, Bernh ...
-
visa fler...
-
Sabatier, Pierre
-
Zubarev, Roman A ...
-
Linder, Stig, 19 ...
-
D´arcy, Pádraig, ...
-
visa färre...
- Om ämnet
-
- MEDICIN OCH HÄLSOVETENSKAP
-
MEDICIN OCH HÄLS ...
-
och Klinisk medicin
-
och Cancer och onkol ...
- Artiklar i publikationen
-
Frontiers in Onc ...
- Av lärosätet
-
Linköpings universitet
-
Karolinska Institutet