The Protein Journal

, Volume 26, Issue 2, pp 125–133 | Cite as

Use of 5′-[p-(Fluorosulfonyl)benzoyl] Guanosine as an Affinity Probe for the Guanine Nucleotide-Binding Site of Transducin



Transducin (T) mediates vision in retinal rods by transmitting light signals detected by rhodopsin to a cGMP phosphodiesterase. The flow of information relies on a subunit association/dissociation cycle of T regulated by a guanine nucleotide exchange/hydrolysis reaction. 5′-[p-(Fluorosulfonyl)benzoyl] guanosine (FSBG) was synthesized and examined here as an affinity label for the guanine nucleotide binding site of T. Although the relative binding affinity of FSBG to T was much lower than for GTP and β,γ-imido-guanosine 5′-triphosphate (GMPPNP), the incorporation of FSBG to T inhibited its light-dependent [3H] GMPPNP binding activity in a concentration dependent manner. Additionally, GDP, GTP and GTP analogs hindered the binding of [3H] FSBG to T. These results demonstrated that FSBG could be used to specifically modify the active site of T. In addition, FSBG was not capable of dissociating T from T:photoactivated rhodopsin complexes, suggesting that in this case FSBG is acting as a GDP analog.


Affinity labeling FSBG G-protein-coupled signaling transducin visual process 



5′-[p-(fluorosulfonyl)benzoyl] guanosine


β,γ-imido-guanosine 5′-triphosphate


G-protein-coupled receptor


guanosine 5′-O-(3-thiotriphosphate)




photoactivated rhodopsin or metarhodopsin II


rod outer segments


sodium dodecyl sulfate


polyacrylamide gel electrophoresis in the presence of SDS




transducin α-subunit


transducin βγ-complex


transducin-photoactivated rhodopsin complex



This work was supported by grants from FONACIT, N° S1-2000000514, Caracas, Venezuela, and from Decanato de Investigación y Desarrollo, N° S1-CB-241, Universidad Simón Bolívar, Caracas, Venezuela. We would like to thank Dr. Graciela L. Uzcanga for helping with the illustrations.


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Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  1. 1.Departamento de QuímicaUniversidad Simón BolívarCaracasVenezuela
  2. 2.Departamento de Biología CelularUniversidad Simón BolívarCaracasVenezuela

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