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KMID : 0390320120220010097
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Chungbuk Medical Journal
2012 Volume.22 No. 1 p.97 ~ p.102
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Correlation between Rho GTPase and Myosin II Activation
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Kim Eung-Gook
Shin Eun-Young
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Abstract
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Purpose: This study analyzed the interaction of myosin II and guanine nucleotide exchange factor (GEF),
PIX, when Rho GTPase Rac was activated by nerve growth factor (NGF) in PC12 cells.
Materials and Methods: PC12 cells were cultured in DMEM with 10% FBS. Cells were starved in
serum-free media for 24 hr, stimulated with 20 ng/ml NGF for indicated times and extracted with cell
lysis buffer to obtain cell lysates. To analyze Rac activation, cell lysates were carried on PBD-His
pulldown assay. Also, to study interaction between PIX and myosin II, cell lysates were
immunoprecipitated with anti-PIX antibody and immunoblotted with anti-PIX or myosin II antibody.
Results: Rac activation showed to increase from 5 min to 20 min by NGF treatment. Interaction of PIX
and myosin was inhibited by NGF from 10 min and dramatically decreased at 20-30 min after NGF
treatment.
Conclusion: These results suggest that NGF-induced Rac activation and dissociation between PIX and
myosin II are correlated with NGF treated time sequences. Therefore, PIX was able to activate Rac
when PIX was separated from myosin II by NGF
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KEYWORD
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PIX, myosin II, Rac
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