Structural Coupling of SH2-Kinase Domains Links Fes and Abl Substrate Recognition and Kinase Activation

Filippakopoulos, P.; Kofler, Michael; Hantschel, Oliver; Gish, Gerald; Grebien, Florian; Salah, E.; Neudecker, Philipp; Kay, Lewis E.; Turk, Benjamin E.; Superti‐Furga, Giulio; Pawson, Tony; Knapp, Stefan

doi:10.1016/j.cell.2008.07.047

Cited by 189 publications

(224 citation statements)

References 53 publications

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“…This conformation has been shown by crystallography to exist in other kinases with SH2-kinase domain arrangements, such as the c-Fes tyrosine kinase (24). Mutagenesis data show that formation of this conformation is essential for maintenance of kinase activity (3,24). This ''tophat'' conformation is perhaps a general feature of the activated conformation of some tyrosine kinases, an idea supported by our HX MS measurements in solution.…”

Section: Resultssupporting

confidence: 62%

“…Taken together, the data comparing Abl(Myr) with Abl(NonMyr) support the conformation in which the SH2 domain occupies a position on top of the small lobe of the kinase domain when the kinase is in the active state. This conformation has been shown by crystallography to exist in other kinases with SH2-kinase domain arrangements, such as the c-Fes tyrosine kinase (24). Mutagenesis data show that formation of this conformation is essential for maintenance of kinase activity (3,24).…”

Section: Resultsmentioning

confidence: 93%

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Conformational disturbance in Abl kinase upon mutation and deregulation

Iacob¹,

Pene-Dumitrescu

Zhang

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

100

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Protein dynamics are inextricably linked to protein function but there are few techniques that allow protein dynamics to be conveniently interrogated. For example, mutations and translocations give rise to aberrant proteins such as Bcr-Abl where changes in protein conformation and dynamics are believed to result in deregulated kinase activity that provides the oncogenic signal in chronic myelogeous leukemia. Although crystal structures of the down-regulated c-Abl kinase core have been reported, the conformational impact of mutations that render Abl resistant to smallmolecule kinase inhibitors are largely unknown as is the allosteric interplay of the various regulatory elements of the protein. Hydrogen exchange mass spectrometry (HX MS) was used to compare the conformations of wild-type Abl with a nonmyristoylated form and with 3 clinically relevant imatinib resistance mutants (T315I, Y253H and E255V). A HX-resistant core localized to the interface between the SH2 and kinase domains, a region known to be important for maintaining the down-regulated state. Conformational differences upon demyristoylation were consistent with the SH2 domain moving to the top of the small lobe of the kinase domain as a function of activation. There were conformational changes in the T315I mutant but, surprisingly, no major changes in conformation were detected in either the Y253H or the E255V mutants. Taken together, these results provide evidence that allosteric interactions and conformational changes play a major role in Abl kinase regulation in solution. Similar analyses could be performed on any protein to provide mechanistic details about conformational changes and protein function. allosteric interactions ͉ chronic mylogenous leukemia ͉ hydrogen exchange ͉ mass spectrometry

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Section: Resultssupporting

confidence: 62%

Section: Resultsmentioning

confidence: 93%

Conformational disturbance in Abl kinase upon mutation and deregulation

Iacob¹,

Pene-Dumitrescu

Zhang

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

100

View full text Add to dashboard Cite

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“…Btk and other Tec family members resemble the Src and Abl families of non-receptor tyrosine kinases in that they contain Src homology 2 and 3 (SH2 and SH3) domains and a kinase domain ( Figure 1B). The SH3-SH2 module plays a multifaceted role of suppressing the kinase before activation, localizing the protein to its target, and stabilizing the kinase after activation (Filippakopoulos et al, 2008;Nagar et al, 2003;Sicheri et al, 1997;Wang et al, 2015;Xu et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

Switch-like activation of Bruton’s tyrosine kinase by membrane-mediated dimerization

Chung

Nocka

Wang

et al. 2018

Preprint

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The transformation of molecular binding events into cellular decisions is the basis of most biological signal transduction. A fundamental challenge faced by these systems is that protein-ligand chemical affinities alone generally result in poor sensitivity to ligand concentration, endangering the system to error. Here, we examine the lipid-binding pleckstrin homology and Tec homology (PH-TH) module of Bruton's tyrosine kinase (Btk) Using fluorescence correlation spectroscopy (FCS) and membrane-binding kinetic measurements, we identify a self-contained phosphatidylinositol (3,4,5)-trisphosphate (PIP 3 ) sensing mechanism that achieves switch-like sensitivity to PIP 3 levels, surpassing the intrinsic affinity discrimination of PIP 3 :PH binding. This mechanism employs multiple PIP 3 binding as well as dimerization of Btk on the membrane surface. Mutational studies in live cells confirm that this mechanism is critical for activation of Btk in vivo. These results demonstrate how a single protein module can institute a minimalist coincidence detection mechanism to achieve high-precision discrimination of ligand concentration.All rights reserved. No reuse allowed without permission.

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“…1B). Consequently, a mutation that disrupts this interface, I164E, inhibits enzyme activity and Bcr-Abl-mediated oncogenesis (8,9). These results suggest that the SH2-kinase interface is a potentially "druggable" site for allosteric inhibition of the kinase activity.…”

mentioning

confidence: 79%

Allosteric Inhibition of Bcr-Abl Kinase by High Affinity Monobody Inhibitors Directed to the Src Homology 2 (SH2)-Kinase Interface

Wojcik

Lamontanara

Grabe

et al. 2016

Journal of Biological Chemistry

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Bcr-Abl is a constitutively active kinase that causes chronic myelogenous leukemia. We have shown that a tandem fusion of two designed binding proteins, termed monobodies, directed to the interaction interface between the Src homology 2 (SH2) and kinase domains and to the phosphotyrosine-binding site of the SH2 domain, respectively, inhibits the Bcr-Abl kinase activity. Because the latter monobody inhibits processive phosphorylation by Bcr-Abl and the SH2-kinase interface is occluded in the active kinase, it remained undetermined whether targeting the SH2-kinase interface alone was sufficient for Bcr-Abl inhibition. To address this question, we generated new, higher affinity monobodies with single nanomolar K D values targeting the kinase-binding surface of SH2. Structural and mutagenesis studies revealed the molecular underpinnings of the monobody-SH2 interactions. Importantly, the new monobodies inhibited Bcr-Abl kinase activity in vitro and in cells, and they potently induced cell death in chronic myelogenous leukemia cell lines. This work provides strong evidence for the SH2-kinase interface as a pharmacologically tractable site for allosteric inhibition of Bcr-Abl.

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Structural Coupling of SH2-Kinase Domains Links Fes and Abl Substrate Recognition and Kinase Activation

Cited by 189 publications

References 53 publications

Conformational disturbance in Abl kinase upon mutation and deregulation

Conformational disturbance in Abl kinase upon mutation and deregulation

Switch-like activation of Bruton’s tyrosine kinase by membrane-mediated dimerization

Allosteric Inhibition of Bcr-Abl Kinase by High Affinity Monobody Inhibitors Directed to the Src Homology 2 (SH2)-Kinase Interface

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