Investigation of the specificity and mechanism of action of the ULK1/AMPK inhibitor SBI-0206965

Research output: Contribution to journalJournal articlepeer-review

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Investigation of the specificity and mechanism of action of the ULK1/AMPK inhibitor SBI-0206965. / Ahwazi, Danial; Neopane, Katyayanee; Markby, Greg R.; Kopietz, Franziska; Ovens, Ashley J; Dall, Morten; Hassing, Anna S; Gräsle, Pamina; Alshuweishi, Yazeed; Treebak, Jonas T.; Salt, Ian P; Göransson, Olga; Zeqiraj, Elton; Scott, John W; Sakamoto, Kei.

In: Biochemical Journal, Vol. 478, No. 15, 2021, p. 2977-2997.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Ahwazi, D, Neopane, K, Markby, GR, Kopietz, F, Ovens, AJ, Dall, M, Hassing, AS, Gräsle, P, Alshuweishi, Y, Treebak, JT, Salt, IP, Göransson, O, Zeqiraj, E, Scott, JW & Sakamoto, K 2021, 'Investigation of the specificity and mechanism of action of the ULK1/AMPK inhibitor SBI-0206965', Biochemical Journal, vol. 478, no. 15, pp. 2977-2997. https://doi.org/10.1042/BCJ20210284

APA

Ahwazi, D., Neopane, K., Markby, G. R., Kopietz, F., Ovens, A. J., Dall, M., Hassing, A. S., Gräsle, P., Alshuweishi, Y., Treebak, J. T., Salt, I. P., Göransson, O., Zeqiraj, E., Scott, J. W., & Sakamoto, K. (2021). Investigation of the specificity and mechanism of action of the ULK1/AMPK inhibitor SBI-0206965. Biochemical Journal, 478(15), 2977-2997. https://doi.org/10.1042/BCJ20210284

Vancouver

Ahwazi D, Neopane K, Markby GR, Kopietz F, Ovens AJ, Dall M et al. Investigation of the specificity and mechanism of action of the ULK1/AMPK inhibitor SBI-0206965. Biochemical Journal. 2021;478(15):2977-2997. https://doi.org/10.1042/BCJ20210284

Author

Ahwazi, Danial ; Neopane, Katyayanee ; Markby, Greg R. ; Kopietz, Franziska ; Ovens, Ashley J ; Dall, Morten ; Hassing, Anna S ; Gräsle, Pamina ; Alshuweishi, Yazeed ; Treebak, Jonas T. ; Salt, Ian P ; Göransson, Olga ; Zeqiraj, Elton ; Scott, John W ; Sakamoto, Kei. / Investigation of the specificity and mechanism of action of the ULK1/AMPK inhibitor SBI-0206965. In: Biochemical Journal. 2021 ; Vol. 478, No. 15. pp. 2977-2997.

Bibtex

@article{5b1d6681a1fe4c20b248e4032dfa2a5f,
title = "Investigation of the specificity and mechanism of action of the ULK1/AMPK inhibitor SBI-0206965",
abstract = "SBI-0206965, originally identified as an inhibitor of the autophagy initiator kinase ULK1, has recently been reported as a more potent and selective AMPK inhibitor relative to the widely used, but promiscuous inhibitor Compound C/Dorsomorphin. Here, we studied the effects of SBI-0206965 on AMPK signalling and metabolic readouts in multiple cell types, including hepatocytes, skeletal muscle cells and adipocytes. We observed SBI-0206965 dose dependently attenuated AMPK-activator (991)-stimulated ACC phosphorylation and inhibition of lipogenesis in hepatocytes. SBI-0206965 (≥ 25 μM) modestly inhibited AMPK signalling in C2C12 myotubes, but also inhibited insulin signalling, insulin-mediated/AMPK-independent glucose uptake, and AICA-riboside uptake. We performed an extended screen of SBI-0206965 against a panel of 140 human protein kinases in vitro, which showed SBI-0206965 inhibits several kinases, including members of AMPK-related kinases (NUAK1, MARK3/4), equally or more potently than AMPK or ULK1. This screen, together with molecular modelling, revealed that most SBI-0206965-sensitive kinases contain a large gatekeeper residue with a preference for methionine at this position. We observed that mutation of the gatekeeper methionine to a smaller side chain amino acid (threonine) rendered AMPK and ULK1 resistant to SBI-0206965 inhibition. These results demonstrate that although SBI-0206965 has utility for delineating AMPK or ULK1 signalling and cellular functions, the compound potently inhibits several other kinases and critical cellular functions such as glucose and nucleoside uptake. Our study demonstrates a role for the gatekeeper residue as a determinant of the inhibitor sensitivity and inhibitor-resistant mutant forms could be exploited as potential controls to probe specific cellular effects of SBI-0206965.",
author = "Danial Ahwazi and Katyayanee Neopane and Markby, {Greg R.} and Franziska Kopietz and Ovens, {Ashley J} and Morten Dall and Hassing, {Anna S} and Pamina Gr{\"a}sle and Yazeed Alshuweishi and Treebak, {Jonas T.} and Salt, {Ian P} and Olga G{\"o}ransson and Elton Zeqiraj and Scott, {John W} and Kei Sakamoto",
note = "Copyright 2021 The Author(s).",
year = "2021",
doi = "10.1042/BCJ20210284",
language = "English",
volume = "478",
pages = "2977--2997",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "15",

}

RIS

TY - JOUR

T1 - Investigation of the specificity and mechanism of action of the ULK1/AMPK inhibitor SBI-0206965

AU - Ahwazi, Danial

AU - Neopane, Katyayanee

AU - Markby, Greg R.

AU - Kopietz, Franziska

AU - Ovens, Ashley J

AU - Dall, Morten

AU - Hassing, Anna S

AU - Gräsle, Pamina

AU - Alshuweishi, Yazeed

AU - Treebak, Jonas T.

AU - Salt, Ian P

AU - Göransson, Olga

AU - Zeqiraj, Elton

AU - Scott, John W

AU - Sakamoto, Kei

N1 - Copyright 2021 The Author(s).

PY - 2021

Y1 - 2021

N2 - SBI-0206965, originally identified as an inhibitor of the autophagy initiator kinase ULK1, has recently been reported as a more potent and selective AMPK inhibitor relative to the widely used, but promiscuous inhibitor Compound C/Dorsomorphin. Here, we studied the effects of SBI-0206965 on AMPK signalling and metabolic readouts in multiple cell types, including hepatocytes, skeletal muscle cells and adipocytes. We observed SBI-0206965 dose dependently attenuated AMPK-activator (991)-stimulated ACC phosphorylation and inhibition of lipogenesis in hepatocytes. SBI-0206965 (≥ 25 μM) modestly inhibited AMPK signalling in C2C12 myotubes, but also inhibited insulin signalling, insulin-mediated/AMPK-independent glucose uptake, and AICA-riboside uptake. We performed an extended screen of SBI-0206965 against a panel of 140 human protein kinases in vitro, which showed SBI-0206965 inhibits several kinases, including members of AMPK-related kinases (NUAK1, MARK3/4), equally or more potently than AMPK or ULK1. This screen, together with molecular modelling, revealed that most SBI-0206965-sensitive kinases contain a large gatekeeper residue with a preference for methionine at this position. We observed that mutation of the gatekeeper methionine to a smaller side chain amino acid (threonine) rendered AMPK and ULK1 resistant to SBI-0206965 inhibition. These results demonstrate that although SBI-0206965 has utility for delineating AMPK or ULK1 signalling and cellular functions, the compound potently inhibits several other kinases and critical cellular functions such as glucose and nucleoside uptake. Our study demonstrates a role for the gatekeeper residue as a determinant of the inhibitor sensitivity and inhibitor-resistant mutant forms could be exploited as potential controls to probe specific cellular effects of SBI-0206965.

AB - SBI-0206965, originally identified as an inhibitor of the autophagy initiator kinase ULK1, has recently been reported as a more potent and selective AMPK inhibitor relative to the widely used, but promiscuous inhibitor Compound C/Dorsomorphin. Here, we studied the effects of SBI-0206965 on AMPK signalling and metabolic readouts in multiple cell types, including hepatocytes, skeletal muscle cells and adipocytes. We observed SBI-0206965 dose dependently attenuated AMPK-activator (991)-stimulated ACC phosphorylation and inhibition of lipogenesis in hepatocytes. SBI-0206965 (≥ 25 μM) modestly inhibited AMPK signalling in C2C12 myotubes, but also inhibited insulin signalling, insulin-mediated/AMPK-independent glucose uptake, and AICA-riboside uptake. We performed an extended screen of SBI-0206965 against a panel of 140 human protein kinases in vitro, which showed SBI-0206965 inhibits several kinases, including members of AMPK-related kinases (NUAK1, MARK3/4), equally or more potently than AMPK or ULK1. This screen, together with molecular modelling, revealed that most SBI-0206965-sensitive kinases contain a large gatekeeper residue with a preference for methionine at this position. We observed that mutation of the gatekeeper methionine to a smaller side chain amino acid (threonine) rendered AMPK and ULK1 resistant to SBI-0206965 inhibition. These results demonstrate that although SBI-0206965 has utility for delineating AMPK or ULK1 signalling and cellular functions, the compound potently inhibits several other kinases and critical cellular functions such as glucose and nucleoside uptake. Our study demonstrates a role for the gatekeeper residue as a determinant of the inhibitor sensitivity and inhibitor-resistant mutant forms could be exploited as potential controls to probe specific cellular effects of SBI-0206965.

U2 - 10.1042/BCJ20210284

DO - 10.1042/BCJ20210284

M3 - Journal article

C2 - 34259310

VL - 478

SP - 2977

EP - 2997

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 15

ER -

ID: 275768073