NDLI: T cell specific adaptor protein (TSAd) promotes interaction of Nck with Lck and SLP-76 in T cells

Content Provider	Springer Nature : BioMed Central
Author	Hem, Cecilie Dahl Sundvold-Gjerstad, Vibeke Granum, Stine Koll, Lise Abrahamsen, Greger Buday, Laszlo Spurkland, Anne
Abstract	Background The Lck and Src binding adaptor protein TSAd (T cell specific adaptor) regulates actin polymerization in T cells and endothelial cells. The molecular details as to how TSAd regulates this process remain to be elucidated. Results To identify novel interaction partners for TSAd, we used a scoring matrix-assisted ligand algorithm (SMALI), and found that the Src homology 2 (SH2) domain of the actin regulator Non-catalytic region of tyrosine kinase adaptor protein (Nck) potentially binds to TSAd phosphorylated on Tyr280 (pTyr280) and pTyr305. These predictions were confirmed by peptide array analysis, showing direct binding of recombinant Nck SH2 to both pTyr280 and pTyr305 on TSAd. In addition, the SH3 domains of Nck interacted with the proline rich region (PRR) of TSAd. Pull-down and immunoprecipitation experiments further confirmed the Nck-TSAd interactions through Nck SH2 and SH3 domains. In line with this Nck and TSAd co-localized in Jurkat cells as assessed by confocal microscopy and imaging flow cytometry. Co-immunoprecipitation experiments in Jurkat TAg cells lacking TSAd revealed that TSAd promotes interaction of Nck with Lck and SLP-76, but not Vav1. TSAd expressing Jurkat cells contained more polymerized actin, an effect dependent on TSAd exon 7, which includes interactions sites for both Nck and Lck. Conclusions TSAd binds to and co-localizes with Nck. Expression of TSAd increases both Nck-Lck and Nck-SLP-76 interaction in T cells. Recruitment of Lck and SLP-76 to Nck by TSAd could be one mechanism by which TSAd promotes actin polymerization in activated T cells.
Related Links	https://biosignaling.biomedcentral.com/counter/pdf/10.1186/s12964-015-0109-7.pdf
Ending Page	14
Page Count	14
Starting Page	1
File Format	HTM / HTML
DOI	10.1186/s12964-015-0109-7
Journal	Cell Communication and Signaling
Issue Number	1
Volume Number	13
Language	English
Publisher	BioMed Central
Publisher Date	2015-07-11
Access Restriction	Open
Subject Keyword	Cell Biology Protein-Ligand Interactions Receptors Cytokines and Growth Factors TSAd T cell specific adaptor protein Nck Lck SH2 domain adaptor protein SH2D2A SLP-76
Content Type	Text
Resource Type	Article
Subject	Biochemistry Cell Biology Molecular Biology
Journal Impact Factor	8.2/2023
5-Year Journal Impact Factor	8/2023

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RESEARCH Open Access T cell specific adaptor protein (TSAd) promotes interaction of Nck with Lck and SLP-76 in T cells

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RESEARCH Open Access T cell specific adaptor protein (TSAd) promotes interaction of Nck with Lck and SLP-76 in T cells

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Tyrosine 192 within the SH2 domain of the Src-protein tyrosine kinase p56Lck regulates T-cell activation independently of Lck/CD45 interactions

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