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PMID: 17967873 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Spatial and temporal regulation of focal adhesion kinase activity in living cells.

Molecular and cellular biology ·Vol. 28 ·No. 1 ·2008-01-00 ·Pages 201-14

Cai X, Lietha D, Ceccarelli DF, Karginov AV, Rajfur Z, Jacobson K, Hahn KM, Eck MJ, Schaller MD

Abstract

Focal adhesion kinase (FAK) is an essential kinase that regulates developmental processes and functions in the pathology of human disease. An intramolecular autoinhibitory interaction between the FERM and catalytic domains is a major mechanism of regulation. Based upon structural studies, a fluorescence resonance energy transfer (FRET)-based FAK biosensor that discriminates between autoinhibited and active conformations of the kinase was developed. This biosensor was used to probe FAK conformational change in live cells and the mechanism of regulation. The biosensor demonstrates directly that FAK undergoes conformational change in vivo in response to activating stimuli. A conserved FERM domain basic patch is required for this conformational change and for interaction with a novel ligand for FAK, acidic phospholipids. Binding to phosphatidylinositol 4,5-bisphosphate (PIP2)-containing phospholipid vesicles activated and induced conformational change in FAK in vitro, and alteration of PIP2 levels in vivo changed the level of activation of the conformational biosensor. These findings provide direct evidence of conformational regulation of FAK in living cells and novel insight into the mechanism regulating FAK conformation.

MeSH Terms
Acids Amino Acid Motifs Amino Acid Sequence Biosensing Techniques Cell Line Cell Survival Focal Adhesion Protein-Tyrosine Kinases/chemistry,genetics,metabolism Humans Models, Molecular Phosphatidylinositol 4,5-Diphosphate/metabolism Phospholipids/metabolism Phosphorylation Protein Binding Protein Structure, Tertiary
Chemicals
Acids Phosphatidylinositol 4,5-Diphosphate Phospholipids Focal Adhesion Protein-Tyrosine Kinases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Cai Xinming
Department of Cell & Developmental Biology, 534 Taylor Hall, CB 7090, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Lietha Daniel
Ceccarelli Derek F
Karginov Andrei V
Rajfur Zenon
Jacobson Ken
Hahn Klaus M
Eck Michael J
Schaller Michael D
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
1098-5549
DOI
10.1128/MCB.01324-07
Published
2008-01-00
Epub
2007-00-29
Pages
201-14
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC2223290
Subset
IM
Grants
NHLBI NIH HHS · P01 HL045100 · United States
NIGMS NIH HHS · U54 GM064346 · United States
NIGMS NIH HHS · GM0064346 · United States
NHLBI NIH HHS · HL45100 · United States
External Links
PubMed source DOI: 10.1128/MCB.01324-07 PMC Full Text View journal details
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