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Mutational Analysis of the Effector Domain of Brassica Sar1 Protein  

Kim, Min-Gab (Environmental Biotechnology National Core Research Center, Gyeongsang National University)
Lee, Jung-Ro (Environmental Biotechnology National Core Research Center, Gyeongsang National University)
Lim, Hye-Song (Environmental Biotechnology National Core Research Center, Gyeongsang National University)
Shin, Mi-Rim (Environmental Biotechnology National Core Research Center, Gyeongsang National University)
Cheon, Min-Gyeong (Environmental Biotechnology National Core Research Center, Gyeongsang National University)
Lee, Deok-Ho (Environmental Biotechnology National Core Research Center, Gyeongsang National University)
Kim, Woe-Yeon (Environmental Biotechnology National Core Research Center, Gyeongsang National University)
Lee, Sang-Yeol (Environmental Biotechnology National Core Research Center, Gyeongsang National University)
Publication Information
Journal of Applied Biological Chemistry / v.50, no.3, 2007 , pp. 109-114 More about this Journal
Abstract
Sar1p is a ras-related GTP-binding protein that functions in intracellular protein transport between the endoplasmic reticulum (ER) and the Golgi complex. The effector domain of Ras family proteins is highly conserved and this domain is functionally interchangeable in plant, yeast and mammalian Sar1. Using a recombinant Brassica sar1 protein (Bsar1p) harboring point mutations in its effector domain, we here investigated the ability of Sar1p to bind and hydrolyze GTP and to interact with the two sar1-specific regulators, GTPase activating protein (GAP) and guanine exchange factor (GEF). The T51A and T55A mutations impaired Bsar1p intrinsic GTP-binding and GDP-dissociation activity. In contrast, mutations in the switch domain of Bsar1 did not affect its intrinsic GTPase activity. Moreover, the P50A, P54A, and S56A mutations affected the interaction between Bsar1p and GAP. P54A mutant protein did not interact with two regulating proteins, GEF and GAP, even though the mutation didn't affect the intrinsic GTP-binding, nucleotide exchange or GTPase activity of Bsar1p.
Keywords
effector domain; GAP; GEF; Mutagenesis; Sar1 GTPase;
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