BioID-Screening Identifies PEAK1 and SHP2 as Components of the ALK Proximitome in Neuroblastoma Cells
Uçkun, Ezgi ; Siaw, Joachim T. LU
; Guan, Jikui
; Anthonydhason, Vimala
; Fuchs, Johannes
; Wolfstetter, Georg
; Hallberg, Bengt
and Palmer, Ruth H.
(2021)
In Journal of Molecular Biology
433(19).
p.1-20
- Abstract
Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase (RTK) that is mutated in approximately 10% of pediatric neuroblastoma (NB). To shed light on ALK-driven signaling processes, we employed BioID-based in vivo proximity labeling to identify molecules that interact intracellularly with ALK. NB-derived SK-N-AS and SK-N-BE(2) cells expressing inducible ALK-BirA* fusion proteins were generated and stimulated with ALKAL ligands in the presence and absence of the ALK tyrosine kinase inhibitor (TKI) lorlatinib. LC/MS-MS analysis identified multiple proteins, including PEAK1 and SHP2, which were validated as ALK interactors in NB cells. Further analysis of the ALK-SHP2 interaction confirmed that the ALK-SHP2 interaction as well as... (More)
Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase (RTK) that is mutated in approximately 10% of pediatric neuroblastoma (NB). To shed light on ALK-driven signaling processes, we employed BioID-based in vivo proximity labeling to identify molecules that interact intracellularly with ALK. NB-derived SK-N-AS and SK-N-BE(2) cells expressing inducible ALK-BirA* fusion proteins were generated and stimulated with ALKAL ligands in the presence and absence of the ALK tyrosine kinase inhibitor (TKI) lorlatinib. LC/MS-MS analysis identified multiple proteins, including PEAK1 and SHP2, which were validated as ALK interactors in NB cells. Further analysis of the ALK-SHP2 interaction confirmed that the ALK-SHP2 interaction as well as SHP2-Y542 phosphorylation was dependent on ALK activation. Use of the SHP2 inhibitors, SHP099 and RMC-4550, resulted in inhibition of cell growth in ALK-driven NB cells. In addition, we noted a strong synergistic effect of combined ALK and SHP2 inhibition that was specific to ALK-driven NB cells, suggesting a potential therapeutic option for ALK-driven NB.
(Less)
- author
- Uçkun, Ezgi
; Siaw, Joachim T.
LU
; Guan, Jikui
; Anthonydhason, Vimala
; Fuchs, Johannes
; Wolfstetter, Georg
; Hallberg, Bengt
and Palmer, Ruth H.
- publishing date
- 2021-09-17
- type
- Contribution to journal
- publication status
- published
- keywords
- BirA*, lorlatinib, RMC-4550, SHP099, signaling
- in
- Journal of Molecular Biology
- volume
- 433
- issue
- 19
- article number
- 167158
- pages
- 1 - 20
- publisher
- Elsevier
- external identifiers
-
- pmid:34273398
- scopus:85111800755
- ISSN
- 0022-2836
- DOI
- 10.1016/j.jmb.2021.167158
- language
- English
- LU publication?
- no
- additional info
- Publisher Copyright: © 2021 The Author(s)
- id
- f1f08297-a825-4cee-ade9-9386979b403a
- date added to LUP
- 2025-03-19 11:49:25
- date last changed
- 2025-03-20 08:02:29
@article{f1f08297-a825-4cee-ade9-9386979b403a,
abstract = {{<p>Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase (RTK) that is mutated in approximately 10% of pediatric neuroblastoma (NB). To shed light on ALK-driven signaling processes, we employed BioID-based in vivo proximity labeling to identify molecules that interact intracellularly with ALK. NB-derived SK-N-AS and SK-N-BE(2) cells expressing inducible ALK-BirA* fusion proteins were generated and stimulated with ALKAL ligands in the presence and absence of the ALK tyrosine kinase inhibitor (TKI) lorlatinib. LC/MS-MS analysis identified multiple proteins, including PEAK1 and SHP2, which were validated as ALK interactors in NB cells. Further analysis of the ALK-SHP2 interaction confirmed that the ALK-SHP2 interaction as well as SHP2-Y542 phosphorylation was dependent on ALK activation. Use of the SHP2 inhibitors, SHP099 and RMC-4550, resulted in inhibition of cell growth in ALK-driven NB cells. In addition, we noted a strong synergistic effect of combined ALK and SHP2 inhibition that was specific to ALK-driven NB cells, suggesting a potential therapeutic option for ALK-driven NB.</p>}},
author = {{Uçkun, Ezgi and Siaw, Joachim T. and Guan, Jikui and Anthonydhason, Vimala and Fuchs, Johannes and Wolfstetter, Georg and Hallberg, Bengt and Palmer, Ruth H.}},
issn = {{0022-2836}},
keywords = {{BirA*; lorlatinib; RMC-4550; SHP099; signaling}},
language = {{eng}},
month = {{09}},
number = {{19}},
pages = {{1--20}},
publisher = {{Elsevier}},
series = {{Journal of Molecular Biology}},
title = {{BioID-Screening Identifies PEAK1 and SHP2 as Components of the ALK Proximitome in Neuroblastoma Cells}},
url = {{http://dx.doi.org/10.1016/j.jmb.2021.167158}},
doi = {{10.1016/j.jmb.2021.167158}},
volume = {{433}},
year = {{2021}},
}