Differences between AGAP1, ASAP1 and Arf GAP1 in substrate recognition: interaction with the N-terminus of Arf1.
Differences between AGAP1, ASAP1 and Arf GAP1 in substrate recognition: interaction with the N-terminus of Arf1.
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AGAP1、ASAP1 和 Arf GAP1 在底物识别方面的差异：与 Arf1 N 末端的相互作用。
DOI：
10.1016/j.cellsig.2004.02.008
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发表时间：
2004
期刊：
影响因子：
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通讯作者：
Randazzo,PaulA
中科院分区：
文献类型：
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作者：
Yoon,Hye-Young;Jacques,Kerry;Nealon,Beth;Stauffer,Stacey;Premont,RichardT;Randazzo,PaulA
Arf间隙是一组结构不同的蛋白质，催化与Arf1结合的GTP的水解。在这里，我们直接比较了Arf1的2-17氨基酸在与三个Arf间隙：ARF Gap1、AGAP1和ASAP1相互作用中的作用。Arf1是一个GTP和磷脂敏感的开关。氨基酸2-17与抗体的隔离抑制了与三个被测试的Arf间隙的相互作用。对Arf1突变体的研究也表明，[2-17]Arf1是与所有三个Arf缺口相互作用的关键结构决定因素；然而，特定突变的影响因缺口而异。与野生型Arf1相比，缺失氨基末端13个氨基酸([Δ13]Arf1)和17个氨基酸([Δ17]Arf1)的Arf1与ASAP1的相互作用分别减少了200和4000倍，与AGAP1的相互作用分别减少了150倍。相反，Arf的氨基末端的缺失使与Arf Gap1的相互作用减少了5倍。通过对点突变的分析，我们发现赖氨酸15和16对Arf1、ASAP1和AGAP1之间的产量相互作用的贡献大于Arf1和Arf Gap1之间的贡献。亮氨酸8参与了与Arf Gap1的相互作用，但不参与与ASAP1和AGAP1的相互作用。Arf1的2-17位氨基酸可抑制Arf Gap1、ASAP1和AGAP1的GAP活性，并直接与ASAP1结合。综上所述，我们的结果表明：(1)Arf Gap与Arf1的2-17个氨基酸相互作用；(2)Arf Gap的每个亚基与Arf1都有一个独特的界面。
The Arf GAPs are a structurally diverse group of proteins that catalyze the hydrolysis of GTP bound to Arf1. Here, we directly compare the role of amino acids 2–17 of Arf1, a GTP- and phospholipid-sensitive switch, for interaction with three Arf GAPs: Arf GAP1, AGAP1 and ASAP1. Sequestration of amino acids 2–17 with an antibody inhibited interaction with the three tested Arf GAPs. Examination of Arf1 mutants also indicated that [2–17]Arf1 is a critical structural determinant of interaction with all three Arf GAPs; however, the effect of specific mutations differed among the GAPs. Compared to wild-type Arf1, Arf1 with the amino terminal 13 ([Δ13]Arf1) and 17 amino acids ([Δ17]Arf1) deleted had 200- and 4000-fold reduced interaction with ASAP1 and 150-fold reduced interaction with AGAP1. In contrast, deletion of the amino terminus of Arf reduced interaction with Arf GAP1 by 5-fold. By analysis of point mutants, we found that lysines 15 and 16 had a greater contribution to productive interaction between Arf1, ASAP1 and AGAP1 than between Arf1 and Arf GAP1. Leucine 8 contributed to the interaction with Arf GAP1 but not with ASAP1 and AGAP1. Amino acids 2–17 of Arf1, isolated from the protein, inhibited GAP activity of Arf GAP1, ASAP1 and AGAP1 and bound directly to ASAP1. Taken together, our results indicate that (i) Arf GAPs interact with amino acids 2–17 of Arf1 and (ii) each subgroup of Arf GAPs has a unique interface with Arf1.