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A novel statin-mediated "prenylation block-and-release" assay provides insight into the membrane targeting mechanisms of small GTPases.

Ali BR, Nouvel I, Leung KF, Hume AN, Seabra MC - Biochem. Biophys. Res. Commun. (2010)

Bottom Line: We found that all Rabs tested were targeted directly to the membrane on which they reside at steady-state and not via an intermediate location as reported for Ras and Rho proteins.However, we observed that the kinetics of cytosol to membrane targeting differed for each Rab tested.Comparison of the mevastatin sensitivity and kinetics of membrane targeting of Rab23, Rab23 prenylation motif mutants and H-Ras revealed that these parameters are strongly dependent upon the prenyl transferase with Rab geranylgeranyl transferase substrates exhibiting higher sensitivity and requiring greater time to recover from mevastatin inhibition than farnesyl transferase substrates.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Faculty of Medicine and Health Sciences, United Arab Emirates University, Al-Ain, United Arab Emirates. bassam.ali@uaeu.ac.ae

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Still images from movies of time-course of Rab membrane targeting following mevastatin prenylation block. HeLa cells expressing EGFP-Rab5a (A) or EGFP-Rab1a (B) in the presence of mevastatin were washed with mevastatin-free medium and images collected at the indicated times (in minutes) following removal of the inhibitor. Images shown in A and B correspond to movies 1 and 2, respectively. Bars = 10 μm.
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d32e579: Still images from movies of time-course of Rab membrane targeting following mevastatin prenylation block. HeLa cells expressing EGFP-Rab5a (A) or EGFP-Rab1a (B) in the presence of mevastatin were washed with mevastatin-free medium and images collected at the indicated times (in minutes) following removal of the inhibitor. Images shown in A and B correspond to movies 1 and 2, respectively. Bars = 10 μm.


A novel statin-mediated "prenylation block-and-release" assay provides insight into the membrane targeting mechanisms of small GTPases.

Ali BR, Nouvel I, Leung KF, Hume AN, Seabra MC - Biochem. Biophys. Res. Commun. (2010)

Still images from movies of time-course of Rab membrane targeting following mevastatin prenylation block. HeLa cells expressing EGFP-Rab5a (A) or EGFP-Rab1a (B) in the presence of mevastatin were washed with mevastatin-free medium and images collected at the indicated times (in minutes) following removal of the inhibitor. Images shown in A and B correspond to movies 1 and 2, respectively. Bars = 10 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2908739&req=5

d32e579: Still images from movies of time-course of Rab membrane targeting following mevastatin prenylation block. HeLa cells expressing EGFP-Rab5a (A) or EGFP-Rab1a (B) in the presence of mevastatin were washed with mevastatin-free medium and images collected at the indicated times (in minutes) following removal of the inhibitor. Images shown in A and B correspond to movies 1 and 2, respectively. Bars = 10 μm.
Bottom Line: We found that all Rabs tested were targeted directly to the membrane on which they reside at steady-state and not via an intermediate location as reported for Ras and Rho proteins.However, we observed that the kinetics of cytosol to membrane targeting differed for each Rab tested.Comparison of the mevastatin sensitivity and kinetics of membrane targeting of Rab23, Rab23 prenylation motif mutants and H-Ras revealed that these parameters are strongly dependent upon the prenyl transferase with Rab geranylgeranyl transferase substrates exhibiting higher sensitivity and requiring greater time to recover from mevastatin inhibition than farnesyl transferase substrates.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Faculty of Medicine and Health Sciences, United Arab Emirates University, Al-Ain, United Arab Emirates. bassam.ali@uaeu.ac.ae

Show MeSH
Related in: MedlinePlus