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

Abstract

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 reliance on protein–ligand chemical affinities alone generally results 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 phosphatidylinositol (3–5)-trisphosphate (PIP 3) sensing mechanism that achieves switch-like sensitivity to PIP 3levels, surpassing the intrinsic affinity discrimination of PIP 3:PH binding. This mechanism employs multiple PIP 3binding as well as dimerization of Btk on the membrane surface. Studies in live cells confirm that mutations at the dimer interface and peripheral site produce effects comparable to that of the kinase-dead Btk in vivo. These results demonstrate how a single protein module can institute an allosteric counting mechanism to achieve high-precision discrimination of ligand concentration. Furthermore, this activation mechanism distinguishes Btk from other Tec family member kinases, Tec and Itk, which we show are not capable of dimerization through their PH-TH modules. This suggests that Btk plays a critical role in the stringency of the B cellmore » response, whereas T cells rely on other mechanisms to achieve stringency.« less

Authors:
 [1];  [1];  [1];  [1];  [2];  [2];  [1];  [3]
  1. Univ. of California, Berkeley, CA (United States)
  2. Univ. of California, San Francisco, CA (United States)
  3. Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC); National Institute of Health
OSTI Identifier:
1561917
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 116; Journal Issue: 22; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Bruton’s tyrosine kinase; PIP3; ultrasensitivity; signaling

Citation Formats

Chung, Jean K., Nocka, Laura M., Decker, Aubrianna, Wang, Qi, Kadlecek, Theresa A., Weiss, Arthur, Kuriyan, John, and Groves, Jay T. Switch-like activation of Bruton’s tyrosine kinase by membrane-mediated dimerization. United States: N. p., 2019. Web. doi:10.1073/pnas.1819309116.
Chung, Jean K., Nocka, Laura M., Decker, Aubrianna, Wang, Qi, Kadlecek, Theresa A., Weiss, Arthur, Kuriyan, John, & Groves, Jay T. Switch-like activation of Bruton’s tyrosine kinase by membrane-mediated dimerization. United States. doi:10.1073/pnas.1819309116.
Chung, Jean K., Nocka, Laura M., Decker, Aubrianna, Wang, Qi, Kadlecek, Theresa A., Weiss, Arthur, Kuriyan, John, and Groves, Jay T. Fri . "Switch-like activation of Bruton’s tyrosine kinase by membrane-mediated dimerization". United States. doi:10.1073/pnas.1819309116.
@article{osti_1561917,
title = {Switch-like activation of Bruton’s tyrosine kinase by membrane-mediated dimerization},
author = {Chung, Jean K. and Nocka, Laura M. and Decker, Aubrianna and Wang, Qi and Kadlecek, Theresa A. and Weiss, Arthur and Kuriyan, John and Groves, Jay T.},
abstractNote = {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 reliance on protein–ligand chemical affinities alone generally results 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 phosphatidylinositol (3–5)-trisphosphate (PIP3) sensing mechanism that achieves switch-like sensitivity to PIP3levels, surpassing the intrinsic affinity discrimination of PIP3:PH binding. This mechanism employs multiple PIP3binding as well as dimerization of Btk on the membrane surface. Studies in live cells confirm that mutations at the dimer interface and peripheral site produce effects comparable to that of the kinase-dead Btk in vivo. These results demonstrate how a single protein module can institute an allosteric counting mechanism to achieve high-precision discrimination of ligand concentration. Furthermore, this activation mechanism distinguishes Btk from other Tec family member kinases, Tec and Itk, which we show are not capable of dimerization through their PH-TH modules. This suggests that Btk plays a critical role in the stringency of the B cell response, whereas T cells rely on other mechanisms to achieve stringency.},
doi = {10.1073/pnas.1819309116},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 22,
volume = 116,
place = {United States},
year = {2019},
month = {5}
}

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