TY - JOUR
T1 - The 14-3-3/SLP76 protein–protein interaction in T-cell receptor signalling
T2 - a structural and biophysical characterization
AU - Soini, Lorenzo
AU - Leysen, Seppe
AU - Davis, Jeremy
AU - Westwood, Marta
AU - Ottmann, Christian
PY - 2021/2
Y1 - 2021/2
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
UR - http://www.scopus.com/inward/record.url?scp=85096700012&partnerID=8YFLogxK
U2 - 10.1002/1873-3468.13993
DO - 10.1002/1873-3468.13993
M3 - Article
C2 - 33159816
AN - SCOPUS:85096700012
VL - 595
SP - 404
EP - 414
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 3
ER -