Summary of FRET-based monitor

Signal transduction monitors based on GFPand FRET

Target

Structure

Ref

Receptors/tyrosine kinases

 

 

 

EGFR

2

EGFR-GFP

anti-PY-Cy3

(1)

 

EGFR

2

EGFP-CFP

Grb2-YFP

(2)

 

Abl/EGF-R

1

YFP-CrkII-CFP (Picchu)

(3)

 

Abl

1

CFP-CrkII-YFP

(4)

 

Insulin R

1

CFP-substrate-SH2-YFP (Phocus)

(5)

 

Src

1

TM(Cbp)-YFP-Cbp(substrate)-SH2 (Csk)-CFP

(6)

 

Src

1

CFP-SH2-substrate-YFP

(4)

 

 

 

 

 

Serine/threonine kinases

 

 

 

 

Raf

1

YFP-Raf-CFP

(7)

 

PKA

1

GFP-KID-BFP

 

(8)

 

PKA

1

CFP-14.3.3-substrate-YFP (AKAR)

(9)

 

PKA

1

RGFP-substrate-BGFP (ART)

(10)

 

AKT/PKB

1

CFP-14.3.3-substrate-YFP [Akt Phocus (Aktus)]

(11)

 

AKT/PKB

1

GFP-AKT-red shifted YFP

(12)

 

PKC

2

PKCa-GFP

phosphospecific Ab-Cy3, 5

(13)

 

PKC

1

CFP-FHA2(Rad53P)-substrate-YFP (CKAR)

(14)

 

 

 

 

 

Small GTPases

 

 

 

 

 

Ras

1

YFP-Ras-Raf-RBD-CFP (Raichu-Ras)

(15)

 

Rap1

1

YFP-Ras-Raf-RBD-CFP (Raichu-Rap1)

(15)

 

Ras

2

Raf-RBD-CFP

CD8-YFP

(16)

 

RalA

1

YFP-RalA-RalBP1-RBD-CFP

(17)

 

RhoA

1

YFP-RhoA-PKN-RBD-CFP (Raichu-Rho)

(18)

 

RhoA

1.5

YFP- RBD-CFP

(18)

 

Rac

1

YFP-PAK-CRIB-Rac-CFP (Raichu-Rac)

(19)

 

Rac

2

GFP-Rac

CRIB-Alexa546

(20)

 

Cdc42

1

YFP-RAK-CRIB-Cdc42-CFP (Raichu-Cdc42)

(19)

 

Rac, Cdc42

1.5

YFP-CRIB-CFP (Raichu-CRIB)

(19)

 

Cdc42

1

YFP-GBD-ACV (N-WASP)-CFP

(21)

 

Cdc42

1.5

YFP-GBD-CFP

(21)

 

Cdc42

1

Cdc42-YFP-GBD-ACV-CFP (GEF sensor)

(21)

 

REFERENCES

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12. Calleja, V., Ameer-Beg, S. M., Vojnovic, B., Woscholski, R., Downward, J., and Larijani, B. (2003) Biochem. J 372, 33-40

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17. Takaya, A., Ohba, Y., Kurokawa, K., and Matsuda, M. (2004) Mol. Biol. Cell 15, 2549-2557

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19. Itoh, R. E., Kurokawa, K., Ohba, Y., Yoshizaki, H., Mochizuki, N., and Matsuda, M. (2002) Mol Cell Biol 22, 6582-6591

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21. Seth, A., Otomo, T., Yin, H. L., and Rosen, M. K. (2003) Biochemistry 42, 3997-4008