An allosteric hot spot in the tandem-SH2 domain of ZAP-70 regulates T-cell signaling

Gangopadhyay, Kaustav; Manna, Bharat; Roy, Swarnendu; Kumari, Sunitha R.; Debnath, Olivia; Chowdhury, Subhankar; Ghosh, Amit; Das, Rahul

doi:10.1042/bcj20190879

Cited by 13 publications

(23 citation statements)

References 75 publications

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“…This behavior has been demonstrated for different protein-protein interaction domains. However, despite their critical roles in many important biological pathways, little information is currently available on Src Homology 2 (SH2) domains, for which only little evidence of interdomain allosteric regulations has been reported (Tsutsui et al, 2016;Gangopadhyay et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

Demonstration of Binding Induced Structural Plasticity in a SH2 Domain

Visconti

Toto

Jarvis

et al. 2020

Front. Mol. Biosci.

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SH2 domains are common protein interaction domains able to recognize short aminoacidic sequences presenting a phosphorylated tyrosine (pY). In spite of their fundamental importance for cell physiology there is a lack of information about the mechanism by which these domains recognize and bind their natural ligands. The N-terminal SH2 (N-SH2) domain of PI3K mediates the interaction with different scaffolding proteins and is known to recognize a specific pY-X-X-M consensus sequence. These interactions are at the cross roads of different molecular pathways and play a key role for cell development and division. By combining mutagenesis, chemical kinetics and NMR, here we provide a complete characterization of the interaction between N-SH2 and a peptide mimicking the scaffolding protein Gab2. Our results highlight that N-SH2 is characterized by a remarkable structural plasticity, with the binding reaction being mediated by a diffused structural region and not solely by the residues located in the binding pocket. Furthermore, the analysis of kinetic data allow us to pinpoint an allosteric network involving residues far from the binding pocket involved in specificity. Results are discussed on the light of previous works on the binding properties of SH2 domains.

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Section: Introductionmentioning

confidence: 99%

Demonstration of Binding Induced Structural Plasticity in a SH2 Domain

Visconti

Toto

Jarvis

et al. 2020

Front. Mol. Biosci.

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“…The resulting data are in agreement with previous studies that showed that the C-SH2 of ZAP-70 undergoes a large conformational change upon binding of an ITAM and that it is dynamic in the absence of a bound peptide. 19,20 Conversely, we find that the C-SH2 of Syk is relatively rigid, even in the unbound state, and does not undergo a similar conformational rearrangement upon binding a peptide. The bias of the Syk tSH2 module towards a conformation resembling that found in the ON state of the kinase, even in the absence of ITAM peptides, suggests that the auto-inhibited state of Syk is less stable than that of ZAP-70.…”

Section: Introductionmentioning

confidence: 77%

“…allowing the binding pocket of the N-SH2 to be fully formed. 19 Binding of the second phosphorylated tyrosine in this pocket remodels the dynamics of the C-SH2 domain binding pocket, resulting in a higher affinity for the phosphorylated ITAM. We show that the C-SH2 of ZAP-70 is more flexible in the ITAM-free state and binding of a peptide rigidifies this domain.…”

Section: Sequence Differences Between the Zap-70 And Syk Tsh2 Modules Are Conservedmentioning

confidence: 99%

Differences in the dynamics of the tandem-SH2 modules of the Syk and ZAP-70 tyrosine kinases

Hobbs

Shah

Badroos

et al. 2021

Preprint

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The catalytic activity of Syk-family tyrosine kinases is regulated by a tandem-SH2 module (tSH2 module). In the autoinhibited state, this module adopts a conformation which stabilizes an inactive conformation of the kinase domain. The binding of the tSH2 module to doubly-phosphorylated tyrosine-containing motifs necessitates a conformational change, thereby relieving kinase inhibition and promoting activation. We determined the crystal structure of the isolated tSH2 module of Syk and find, in contrast to ZAP-70, that its conformation more closely resembles that of the peptide-bound state, rather than the autoinhibited state. Hydrogen-deuterium exchange by mass spectrometry, as well as molecular dynamics simulations, reveal that the dynamics of the tSH2 modules of Syk and ZAP-70 differ, with most of these differences occurring in the C-terminal SH2 domain. Our data suggest that the conformational landscapes of the tSH2 modules in Syk and ZAP-70 have been tuned differently, such that the auto-inhibited conformation of the Syk tSH2 module is less stable. This feature of Syk likely contributes to its ability to more readily escape autoinhibition when compared to ZAP-70, consistent with tighter control of downstream signaling pathways in T cells.

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“…Steady-state ligand-binding experiments suggest that the tSH2 domain binds to the doubly phosphorylated ITAM peptides in a biphasic manner [ 179 ]. In the first step, the C-SH2 domain binds uncooperatively to the N-terminal tyrosine phosphate with low nanomolar affinity, subsequently facilitating the formation of the N-SH2 phosphate-binding pocket enabling second phosphotyrosine to bind with micromolar affinity leading to remodeling of C-SH2-binding site.…”

Section: Regulation Of Zap-70 Activationmentioning

confidence: 99%

“…The functional significance of such two-step ligand binding is still not completely understood. Comparison of the apo- and holo- structure of the tSH2 domain revealed that the N-SH2-binding pocket formed at the interface of the tSH2 domains [ 169 , 174 , 179 , 182 ]. A non-covalent network of amino acids residues allosterically couples the tSH2 domains during ligand binding.…”

Section: Regulation Of Zap-70 Activationmentioning

confidence: 99%

Regulating the discriminatory response to antigen by T-cell receptor

et al. 2022

Self Cite

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The cell-mediated immune response constitutes a robust host defense mechanism to eliminate pathogens and oncogenic cells. T-cells play a central role in such a defense mechanism and creating memories to prevent any potential infection. T-cell recognizes foreign antigen by its surface receptors when presented through antigen-presenting cells and calibrates its cellular response by a network of intracellular signaling events. Activation of T-cell receptor (TCR) leads to changes in gene expression and metabolic networks regulating cell development, proliferation, and migration. TCR does not possess any catalytic activity, and the signaling initiates with the colocalization of several enzymes and scaffold proteins. Deregulation of T cell signaling is often linked to autoimmune disorders like SCID, Rheumatoid arthritis, and multiple sclerosis. The TCR remarkably distinguishes the minor difference between self and non-self antigen through a kinetic proofreading mechanism. The output of TCR signaling is determined by the half-life of the receptor-antigen complex and the time taken to recruit and activate the downstream enzymes. A longer half-life of a non-self antigen receptor complex could initiate downstream signaling by activating associated enzymes. Whereas, the short-lived self-peptide receptor complex disassembles before the downstream enzymes are activated. Activation of TCR rewires the cellular metabolic response to aerobic glycolysis from oxidative phosphorylation. How does the early event in the TCR signaling cross-talk with the cellular metabolism is an open question. In this review, we have discussed the recent developments in understanding the regulation of TCR signaling, and then we reviewed the emerging role of metabolism in regulating T-cell function.

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An allosteric hot spot in the tandem-SH2 domain of ZAP-70 regulates T-cell signaling

Cited by 13 publications

References 75 publications

Demonstration of Binding Induced Structural Plasticity in a SH2 Domain

Demonstration of Binding Induced Structural Plasticity in a SH2 Domain

Differences in the dynamics of the tandem-SH2 modules of the Syk and ZAP-70 tyrosine kinases

Regulating the discriminatory response to antigen by T-cell receptor

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