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The proximity-labeling technique BioID identifies sorting nexin 6 as a member of the insulin-like growth factor 1 (IGF1)-IGF1 receptor pathway.

Publication ,  Journal Article
Bareja, A; Hodgkinson, CP; Soderblom, E; Waitt, G; Dzau, VJ
Published in: J Biol Chem
April 27, 2018

The insulin-like growth factor 1 receptor (IGF1R) is a receptor tyrosine kinase with critical roles in various biological processes. Recent results from clinical trials targeting IGF1R indicate that IGF1R signaling pathways are more complex than previously thought. Moreover, it has become increasingly clear that the function of many proteins can be understood only in the context of a network of interactions. To that end, we sought to profile IGF1R-protein interactions with the proximity-labeling technique BioID. We applied BioID by generating a HEK293A cell line that stably expressed the BirA* biotin ligase fused to the IGF1R. Following stimulation by IGF1, biotinylated proteins were analyzed by MS. This screen identified both known and previously unknown interactors of IGF1R. One of the novel interactors was sorting nexin 6 (SNX6), a protein that forms part of the retromer complex, which is involved in intracellular protein sorting. Using co-immunoprecipitation, we confirmed that IGF1R and SNX6 physically interact. SNX6 knockdown resulted in a dramatic diminution of IGF1-mediated ERK1/2 phosphorylation, but did not affect IGF1R internalization. Bioluminescence resonance energy transfer experiments indicated that the SNX6 knockdown perturbed the association between IGF1R and the key adaptor proteins insulin receptor substrate 1 (IRS1) and SHC adaptor protein 1 (SHC1). Intriguingly, even in the absence of stimuli, SNX6 overexpression significantly increased Akt phosphorylation. Our study confirms the utility of proximity-labeling methods, such as BioID, to screen for interactors of cell-surface receptors and has uncovered a role of one of these interactors, SNX6, in the IGF1R signaling cascade.

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Published In

J Biol Chem

DOI

EISSN

1083-351X

Publication Date

April 27, 2018

Volume

293

Issue

17

Start / End Page

6449 / 6459

Location

United States

Related Subject Headings

  • Staining and Labeling
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Sorting Nexins
  • Repressor Proteins
  • Receptors, Somatomedin
  • Receptor, IGF Type 1
  • Proto-Oncogene Proteins c-akt
  • Phosphorylation
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinase 1
 

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Bareja, A., Hodgkinson, C. P., Soderblom, E., Waitt, G., & Dzau, V. J. (2018). The proximity-labeling technique BioID identifies sorting nexin 6 as a member of the insulin-like growth factor 1 (IGF1)-IGF1 receptor pathway. J Biol Chem, 293(17), 6449–6459. https://doi.org/10.1074/jbc.RA118.002406
Bareja, Akshay, Conrad P. Hodgkinson, Erik Soderblom, Greg Waitt, and Victor J. Dzau. “The proximity-labeling technique BioID identifies sorting nexin 6 as a member of the insulin-like growth factor 1 (IGF1)-IGF1 receptor pathway.J Biol Chem 293, no. 17 (April 27, 2018): 6449–59. https://doi.org/10.1074/jbc.RA118.002406.
Bareja A, Hodgkinson CP, Soderblom E, Waitt G, Dzau VJ. The proximity-labeling technique BioID identifies sorting nexin 6 as a member of the insulin-like growth factor 1 (IGF1)-IGF1 receptor pathway. J Biol Chem. 2018 Apr 27;293(17):6449–59.
Bareja, Akshay, et al. “The proximity-labeling technique BioID identifies sorting nexin 6 as a member of the insulin-like growth factor 1 (IGF1)-IGF1 receptor pathway.J Biol Chem, vol. 293, no. 17, Apr. 2018, pp. 6449–59. Pubmed, doi:10.1074/jbc.RA118.002406.
Bareja A, Hodgkinson CP, Soderblom E, Waitt G, Dzau VJ. The proximity-labeling technique BioID identifies sorting nexin 6 as a member of the insulin-like growth factor 1 (IGF1)-IGF1 receptor pathway. J Biol Chem. 2018 Apr 27;293(17):6449–6459.

Published In

J Biol Chem

DOI

EISSN

1083-351X

Publication Date

April 27, 2018

Volume

293

Issue

17

Start / End Page

6449 / 6459

Location

United States

Related Subject Headings

  • Staining and Labeling
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Sorting Nexins
  • Repressor Proteins
  • Receptors, Somatomedin
  • Receptor, IGF Type 1
  • Proto-Oncogene Proteins c-akt
  • Phosphorylation
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinase 1