Archives of Biochemistry and Biophysics 2012-08-15

Characterization of heterotrimeric nucleotide-depleted Gαi-proteins by Bodipy-FL-GTPγS fluorescence anisotropy

Lauri Tõntson, Anna Babina, Taavi Võsumaa, Sergei Kopanchuk, Ago Rinken, Lauri Tõntson, Anna Babina, Taavi Võsumaa, Sergei Kopanchuk, Ago Rinken

Index: Arch. Biochem. Biophys. 524(2) , 93-8, (2012)

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Abstract

Highlights ► StrepII-tagged γ2-subunits for the purification of heterotrimeric G-proteins. ► Preparation of GDP-depleted αi. ► Real-time monitoring of Bodipy-FL-GTPγS binding. ► Instability of GDP-depleted αi at room temperature. ► Mn2+ accelerates nucleotide release from Gα.