The 14-3-3/SLP76 protein–protein interaction in T-cell receptor signalling: a structural and biophysical characterization

Lorenzo Soini; Seppe Leysen; Jeremy Davis; Marta Westwood; Christian Ottmann

doi:10.1002/1873-3468.13993

The 14-3-3/SLP76 protein–protein interaction in T-cell receptor signalling: a structural and biophysical characterization

Lorenzo Soini
, Seppe Leysen
, Jeremy Davis
, Marta Westwood
, Christian Ottmann (Corresponding author)

Research output: Contribution to journal › Article › Academic › peer-review

8 Citations (Scopus)

Abstract

The SH2 domain-containing protein of 76 kDa, SLP76, is an important adaptor protein that coordinates a complex protein network downstream of T-cell receptors (TCR), ultimately regulating the immune response. Upon phosphorylation on Ser376, SLP76 interacts with 14-3-3 adaptor proteins, which leads to its proteolytic degradation. This provides a negative feedback mechanism by which TCR signalling can be controlled. To gain insight into the 14-3-3/SLP76 protein–protein interaction (PPI), we have determined a high-resolution crystal structure of a SLP76 synthetic peptide containing Ser376 with 14-3-3σ. We then characterized its binding to 14-3-3 proteins biophysically by means of fluorescence polarization and isothermal titration calorimetry. Furthermore, we generated two recombinant SLP76 protein constructs and characterized their binding to 14-3-3. Our work lays the foundation for drug design efforts aimed at targeting the 14-3-3/SLP76 interaction and, thereby, TCR signalling.

Original language	English
Pages (from-to)	404-414
Number of pages	11
Journal	FEBS Letters
Volume	595
Issue number	3
DOIs	https://doi.org/10.1002/1873-3468.13993
Publication status	Published - Feb 2021

Funding

This work is part of the TASPPI project. The TASPPI project is supported by the Initial Training Network (ITN) initiative, funded by the H2020 Marie Curie Actions of the European Commission under Grant Agreement 675179. We thank Dr. Rebecca Burnley and Dr. Victoria Ellis for performing the Native MS.

Funders	Funder number
Marie Skłodowska‐Curie
Marie Skłodowska‐Curie
European Commission	675179

Keywords

isothermal titration calorimetry
protein constructs
surface plasmon resonance
TCR signalling and 14-3-3
X-ray protein crystallography

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title = "The 14-3-3/SLP76 protein–protein interaction in T-cell receptor signalling: a structural and biophysical characterization",

abstract = "The SH2 domain-containing protein of 76 kDa, SLP76, is an important adaptor protein that coordinates a complex protein network downstream of T-cell receptors (TCR), ultimately regulating the immune response. Upon phosphorylation on Ser376, SLP76 interacts with 14-3-3 adaptor proteins, which leads to its proteolytic degradation. This provides a negative feedback mechanism by which TCR signalling can be controlled. To gain insight into the 14-3-3/SLP76 protein–protein interaction (PPI), we have determined a high-resolution crystal structure of a SLP76 synthetic peptide containing Ser376 with 14-3-3σ. We then characterized its binding to 14-3-3 proteins biophysically by means of fluorescence polarization and isothermal titration calorimetry. Furthermore, we generated two recombinant SLP76 protein constructs and characterized their binding to 14-3-3. Our work lays the foundation for drug design efforts aimed at targeting the 14-3-3/SLP76 interaction and, thereby, TCR signalling.",

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author = "Lorenzo Soini and Seppe Leysen and Jeremy Davis and Marta Westwood and Christian Ottmann",

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AU - Soini, Lorenzo

AU - Leysen, Seppe

AU - Davis, Jeremy

AU - Westwood, Marta

AU - Ottmann, Christian

PY - 2021/2

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N2 - The SH2 domain-containing protein of 76 kDa, SLP76, is an important adaptor protein that coordinates a complex protein network downstream of T-cell receptors (TCR), ultimately regulating the immune response. Upon phosphorylation on Ser376, SLP76 interacts with 14-3-3 adaptor proteins, which leads to its proteolytic degradation. This provides a negative feedback mechanism by which TCR signalling can be controlled. To gain insight into the 14-3-3/SLP76 protein–protein interaction (PPI), we have determined a high-resolution crystal structure of a SLP76 synthetic peptide containing Ser376 with 14-3-3σ. We then characterized its binding to 14-3-3 proteins biophysically by means of fluorescence polarization and isothermal titration calorimetry. Furthermore, we generated two recombinant SLP76 protein constructs and characterized their binding to 14-3-3. Our work lays the foundation for drug design efforts aimed at targeting the 14-3-3/SLP76 interaction and, thereby, TCR signalling.

AB - The SH2 domain-containing protein of 76 kDa, SLP76, is an important adaptor protein that coordinates a complex protein network downstream of T-cell receptors (TCR), ultimately regulating the immune response. Upon phosphorylation on Ser376, SLP76 interacts with 14-3-3 adaptor proteins, which leads to its proteolytic degradation. This provides a negative feedback mechanism by which TCR signalling can be controlled. To gain insight into the 14-3-3/SLP76 protein–protein interaction (PPI), we have determined a high-resolution crystal structure of a SLP76 synthetic peptide containing Ser376 with 14-3-3σ. We then characterized its binding to 14-3-3 proteins biophysically by means of fluorescence polarization and isothermal titration calorimetry. Furthermore, we generated two recombinant SLP76 protein constructs and characterized their binding to 14-3-3. Our work lays the foundation for drug design efforts aimed at targeting the 14-3-3/SLP76 interaction and, thereby, TCR signalling.

KW - isothermal titration calorimetry

KW - protein constructs

KW - surface plasmon resonance

KW - TCR signalling and 14-3-3

KW - X-ray protein crystallography

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ER -

The 14-3-3/SLP76 protein–protein interaction in T-cell receptor signalling: a structural and biophysical characterization

Abstract

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Keywords

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