Figure 6.

Rad is a regulator of NM23. (A) GST-nm23 (50 ng) was incubated with 3 μCi each of [α-³²P]GTP, [α-³²P]CTP, [α-³²P]TTP, and [α-³²P]ATP in the absence or presence of 1 μg GST-Rad. After 5 min at 25°C, the reaction was stopped with 10-fold cold buffer, and the nucleotides were resolved by PEI-TLC [mobile phase = 0.1 M LiCl and 0.06 M (NH₄)SO₄] and were quantitated by PhosphorImager analysis, and the percent conversion was calculated as the ratio of NDP/(NDP + NTP) × 100. (B) Wild-type and Pro96 GST-nm23 (0.5 μg) was incubated with 5 μCi of [γ-³²P]ATP in the absence or presence of 1 μg of GST-Rad at 25°C for 5 min. After addition of 2 × SDS/PAGE sample buffer (pH 8.8), the mixture was incubated at 25°C for 10 min, and proteins were resolved by 10% SDS/PAGE.