TY - JOUR

T1 - Phosphotyrosine couples peptide binding and SHP2 activation via a dynamic allosteric network

AU - Marasco, Michelangelo

AU - Kirkpatrick, John

AU - Nanna, Vittoria

AU - Sikorska, Justyna

AU - Carlomagno, Teresa

N1 - Funding Information: This work was funded by the German Science Foundation DFG (grant CA 294/20-1 to TC). MM was supported by a fellowship from the Hannover School for Biomolecular Drug Research (HSBDR) and was a member of the Hannover Biomedical Research School (HBRS) and the MD/PhD program “Molecular Medicine”.

PY - 2021

Y1 - 2021

N2 - SHP2 is a ubiquitous protein tyrosine phosphatase, whose activity is regulated by phosphotyrosine (pY)-containing peptides generated in response to extracellular stimuli. Its crystal structure reveals a closed, auto-inhibited conformation in which the N-terminal Src homology 2 (N-SH2) domain occludes the catalytic site of the phosphatase (PTP) domain. High-affinity mono-phosphorylated peptides promote catalytic activity by binding to N-SH2 and disrupting the interaction with the PTP. The mechanism behind this process is not entirely clear, especially because N-SH2 is incapable of accommodating complete peptide binding when SHP2 is in the auto-inhibited state. Here, we show that pY performs an essential role in this process; in addition to its contribution to overall peptide-binding energy, pY-recognition leads to enhanced dynamics of the N-SH2 EF and BG loops via an allosteric communication network, which destabilizes the N-SH2–PTP interaction surface and simultaneously generates a fully accessible binding pocket for the C-terminal half of the phosphopeptide. Subsequently, full binding of the phosphopeptide is associated with the stabilization of activated SHP2. We demonstrate that this allosteric network exists only in N-SH2, which is directly involved in the regulation of SHP2 activity, while the C-terminal SH2 domain (C-SH2) functions primarily to recruit high-affinity bidentate phosphopeptides.

AB - SHP2 is a ubiquitous protein tyrosine phosphatase, whose activity is regulated by phosphotyrosine (pY)-containing peptides generated in response to extracellular stimuli. Its crystal structure reveals a closed, auto-inhibited conformation in which the N-terminal Src homology 2 (N-SH2) domain occludes the catalytic site of the phosphatase (PTP) domain. High-affinity mono-phosphorylated peptides promote catalytic activity by binding to N-SH2 and disrupting the interaction with the PTP. The mechanism behind this process is not entirely clear, especially because N-SH2 is incapable of accommodating complete peptide binding when SHP2 is in the auto-inhibited state. Here, we show that pY performs an essential role in this process; in addition to its contribution to overall peptide-binding energy, pY-recognition leads to enhanced dynamics of the N-SH2 EF and BG loops via an allosteric communication network, which destabilizes the N-SH2–PTP interaction surface and simultaneously generates a fully accessible binding pocket for the C-terminal half of the phosphopeptide. Subsequently, full binding of the phosphopeptide is associated with the stabilization of activated SHP2. We demonstrate that this allosteric network exists only in N-SH2, which is directly involved in the regulation of SHP2 activity, while the C-terminal SH2 domain (C-SH2) functions primarily to recruit high-affinity bidentate phosphopeptides.

KW - Allosteric coupling

KW - Molecular dynamics

KW - NMR spectroscopy

KW - PD-1

KW - SHP2

UR - http://www.scopus.com/inward/record.url?scp=85105096884&partnerID=8YFLogxK

U2 - 10.1016/j.csbj.2021.04.040

DO - 10.1016/j.csbj.2021.04.040

M3 - Article

AN - SCOPUS:85105096884

VL - 19

SP - 2398

EP - 2415

JO - Computational and structural biotechnology journal

JF - Computational and structural biotechnology journal

SN - 2001-0370

ER -