TY  - JOUR
AU  - Möhl, C.
AU  - Kirchgeßner, N.
AU  - Schäfer, C.
AU  - Küpper, K.
AU  - Born, S.
AU  - Diez, G.
AU  - Goldmann, W.H.
AU  - Merkel, R.
AU  - Hoffmann, B.
TI  - Becoming Stable and Strong: The Interplay between Vinculin Exchange Dynamics and Adhesion Strength During Adhesion Site Maturation
JO  - Cell Motility and the Cytoskeleton
VL  - 66
SN  - 0886-1544
CY  - Bognor Regis
PB  - Wiley
M1  - PreJuSER-4910
SP  - 350 - 364
PY  - 2009
N1  - Contract grant sponsors: BFHZ, BaCaTec, and DAAD.
AB  - The coordinated formation and release of focal adhesions is necessary for cell attachment and migration. According to current models, these processes are caused by temporal variations in protein composition. Protein incorporation into focal adhesions is believed to be controlled by phosphorylation. Here, we tested the exchange dynamics of GFP-vinculin as marker protein of focal adhesions using the method of Fluorescence Recovery After Photobleaching. The relevance of the phosphorylation state of the protein, the age of focal adhesions and the acting force were investigated. For stable focal adhesions of stationary keratinocytes, we determined an exchangeable vinculin fraction of 52% and a recovery halftime of 57 s. Nascent focal adhesions of moving cells contained a fraction of exchanging vinculin of 70% with a recovery halftime of 36 s. Upon maturation, mean saturation values and recovery halftimes decreased to levels of 49% and 42 s, respectively. Additionally, the fraction of stably incorporated vinculin increased with cell forces and decreased with vinculin phosphorylation within these sites. Experiments on a nonphosphorylatable vinculin mutant construct at phosphorylation site tyr1065 confirmed the direct interplay between phosphorylation and exchange dynamics of adhesion proteins during adhesion site maturation.
KW  - Cell Adhesion: physiology
KW  - Cell Movement: physiology
KW  - Cells, Cultured
KW  - Fluorescence Recovery After Photobleaching
KW  - Focal Adhesions: metabolism
KW  - Humans
KW  - Keratinocytes: cytology
KW  - Keratinocytes: metabolism
KW  - Phosphorylation: physiology
KW  - Vinculin: genetics
KW  - Vinculin: metabolism
KW  - Vinculin (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:19422016
UR  - <Go to ISI:>//WOS:000266466700005
DO  - DOI:10.1002/cm.20375
UR  - https://juser.fz-juelich.de/record/4910
ER  -