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Title: Structural Basis for the Autoinhibition of Focal Adhesion Kinase

Abstract

Appropriate tyrosine kinase signaling depends on coordinated sequential coupling of protein-protein interactions with catalytic activation. Focal adhesion kinase (FAK) integrates signals from integrin and growth factor receptors to regulate cellular responses including cell adhesion, migration, and survival. Here, we describe crystal structures representing both autoinhibited and active states of FAK. The inactive structure reveals a mechanism of inhibition in which the N-terminal FERM domain directly binds the kinase domain, blocking access to the catalytic cleft and protecting the FAK activation loop from Src phosphorylation. Additionally, the FERM domain sequesters the Tyr397 autophosphorylation and Src recruitment site, which lies in the linker connecting the FERM and kinase domains. The active phosphorylated FAK kinase adopts a conformation that is immune to FERM inhibition. Our biochemical and structural analysis shows how the architecture of autoinhibited FAK orchestrates an activation sequence of FERM domain displacement, linker autophosphorylation, Src recruitment, and full catalytic activation.

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL) National Synchrotron Light Source
Sponsoring Org.:
Doe - Office Of Science
OSTI Identifier:
930438
Report Number(s):
BNL-81183-2008-JA
Journal ID: ISSN 0092-8674; CELLB5; TRN: US200904%%712
DOE Contract Number:
DE-AC02-98CH10886
Resource Type:
Journal Article
Resource Relation:
Journal Name: Cell; Journal Volume: 129
Country of Publication:
United States
Language:
English
Subject:
36 MATERIALS SCIENCE; ADHESION; CRYSTAL STRUCTURE; GROWTH FACTORS; PHOSPHORYLATION; PHOSPHOTRANSFERASES; PROTEINS; RECEPTORS; TYROSINE; national synchrotron light source

Citation Formats

Lietha,D., Cai, X., Ceccarelli, D., Lietha, Y., Schaller, M., and Eck, M. Structural Basis for the Autoinhibition of Focal Adhesion Kinase. United States: N. p., 2007. Web. doi:10.1016/j.cell.2007.05.041.
Lietha,D., Cai, X., Ceccarelli, D., Lietha, Y., Schaller, M., & Eck, M. Structural Basis for the Autoinhibition of Focal Adhesion Kinase. United States. doi:10.1016/j.cell.2007.05.041.
Lietha,D., Cai, X., Ceccarelli, D., Lietha, Y., Schaller, M., and Eck, M. Mon . "Structural Basis for the Autoinhibition of Focal Adhesion Kinase". United States. doi:10.1016/j.cell.2007.05.041.
@article{osti_930438,
title = {Structural Basis for the Autoinhibition of Focal Adhesion Kinase},
author = {Lietha,D. and Cai, X. and Ceccarelli, D. and Lietha, Y. and Schaller, M. and Eck, M.},
abstractNote = {Appropriate tyrosine kinase signaling depends on coordinated sequential coupling of protein-protein interactions with catalytic activation. Focal adhesion kinase (FAK) integrates signals from integrin and growth factor receptors to regulate cellular responses including cell adhesion, migration, and survival. Here, we describe crystal structures representing both autoinhibited and active states of FAK. The inactive structure reveals a mechanism of inhibition in which the N-terminal FERM domain directly binds the kinase domain, blocking access to the catalytic cleft and protecting the FAK activation loop from Src phosphorylation. Additionally, the FERM domain sequesters the Tyr397 autophosphorylation and Src recruitment site, which lies in the linker connecting the FERM and kinase domains. The active phosphorylated FAK kinase adopts a conformation that is immune to FERM inhibition. Our biochemical and structural analysis shows how the architecture of autoinhibited FAK orchestrates an activation sequence of FERM domain displacement, linker autophosphorylation, Src recruitment, and full catalytic activation.},
doi = {10.1016/j.cell.2007.05.041},
journal = {Cell},
number = ,
volume = 129,
place = {United States},
year = {Mon Jan 01 00:00:00 EST 2007},
month = {Mon Jan 01 00:00:00 EST 2007}
}
  • The LIM-only adaptor PINCH (the particularly interesting cysteine- and histidine-rich protein) plays a pivotal role in the assembly of focal adhesions (FAs), supramolecular complexes that transmit mechanical and biochemical information between extracellular matrix and actin cytoskeleton, regulating diverse cell adhesive processes such as cell migration, cell spreading, and survival. A key step for the PINCH function is its localization to FAs, which depends critically on the tight binding of PINCH to integrin-linked kinase (ILK). Here we report the solution NMR structure of the core ILK {center_dot} PINCH complex (28 kDa, K{sub D} {approx} 68 nm) involving the N-terminal ankyrin repeatmore » domain (ARD) of ILK and the first LIM domain (LIM1) of PINCH. We show that the ILK ARD exhibits five sequentially stacked ankyrin repeat units, which provide a large concave surface to grip the two contiguous zinc fingers of the PINCH LIM1. The highly electrostatic interface is evolutionally conserved but differs drastically from those of known ARD and LIM bound to other types of protein domains. Consistently mutation of a hot spot in LIM1, which is not conserved in other LIM domains, disrupted the PINCH binding to ILK and abolished the PINCH targeting to FAs. These data provide atomic insight into a novel modular recognition and demonstrate how PINCH is specifically recruited by ILK to mediate the FA assembly and cell-extracellular matrix communication.« less
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