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In vitro GAP activity of Git2-short toward ARF1. ARF1 (0.5 μg) preloaded with [α-32P]GTP were incubated for 5 min at 30°C with buffer alone (lane 1), with 2.5 μg of Git2-short (lane 2), or the CA mutant (lane 3). The nucleotides were then extracted and separated on a polyethyleneimine-cellulose plate and subjected to autoradiography. Positions of GTP and GDP were identified using authentic nucleotides.
