Finazzi D, Cassel D, Donaldson J G, Klausner R D
Cell Biology and Metabolism Branch, NICHD, Bethesda, Maryland 20892.
J Biol Chem. 1994 May 6;269(18):13325-30.
The GTP-dependent interaction of ADP ribosylation factor 1 (ARF1) with Golgi membranes is required for the binding of cytosolic coatomer proteins to those membranes. Whereas both GTP and GTP gamma S can support coatomer binding to membranes, by using partially purified components, GTP-driven binding is rapidly reversible (t1/2 of 2 min) while that driven by GTP gamma S is more stable (t1/2 of over 30 min). In the presence of GTP, aluminum fluoride, an activator of trimeric G proteins, promotes the stable ARF-dependent binding of coatomer to membranes, even though this reagent does not itself activate ARF. Aluminum fluoride appears to act, like GTP gamma S, to make the binding of coatomer relatively irreversible. It acts to inhibit ARF-GTP hydrolysis catalyzed by the membrane and thus makes the ARF-GTP active state persistent. This effect is not dependent on the presence of any cytosolic component, such as the coatomer. The number of molecules of ARF that can be protected from hydrolysis by aluminum fluoride, however, is only a fraction of the total amount of ARF that can bind to membranes in the presence of GTP gamma S. We propose that this population defines a set of binding sites that are sufficient for coat protein assembly onto the membrane.
ADP核糖基化因子1(ARF1)与高尔基体膜的GTP依赖性相互作用是胞质衣被蛋白结合到这些膜上所必需的。虽然GTP和GTPγS都能支持衣被蛋白与膜的结合,但使用部分纯化的组分时,GTP驱动的结合是快速可逆的(半衰期为2分钟),而GTPγS驱动的结合更稳定(半衰期超过30分钟)。在GTP存在的情况下,三聚体G蛋白的激活剂氟化铝能促进衣被蛋白与膜的稳定的ARF依赖性结合,尽管该试剂本身并不激活ARF。氟化铝似乎像GTPγS一样起作用,使衣被蛋白的结合相对不可逆。它的作用是抑制膜催化的ARF-GTP水解,从而使ARF-GTP活性状态持续存在。这种效应不依赖于任何胞质组分的存在,如衣被蛋白。然而,能被氟化铝保护免于水解的ARF分子数量只是在GTPγS存在下能结合到膜上的ARF总量的一小部分。我们认为,这一群结合位点足以使衣被蛋白组装到膜上。