PMID: 9182698Jun 1, 1997Paper

Resolution of two ADP-ribosylation factor 1 GTPase-activating proteins from rat liver

The Biochemical Journal
Paul A Randazzo

Abstract

ADP-ribosylation factor 1 (ARF1) is a 21 kDa GTP-binding protein that regulates multiple steps in membrane traffic. Here, two ARF1 GTPase-activating proteins (GAPs) from rat liver were resolved. The GAPs were antigenically distinct. One reacted with a polyclonal antibody raised against the GAP catalytic peptide previously purified by Makler et al. [Makler, Cukierman, Rotman, Admon and Cassel (1995) J. Biol. Chem. 270, 5232-5237], and here is referred to as GAP1. The other GAP (GAP2) did not react with the antibody. These GAPs differed in phospholipid dependencies. GAP1 was activated 3-7-fold by the acid phospholipids phosphatidylinositol 4, 5-bisphosphate (PIP2), phosphatidic acid (PA) and phosphatidylserine (PS). In contrast, GAP2 was stimulated 20-40-fold by PIP2. PA and PS had no effect by themselves but PA increased GAP2 activity in the presence of PIP2. The GAPs were otherwise similar in activity. In the presence of phosphoinositides, the Km of GAP1 for ARF1-GTP was estimated to be 8.1+/-1.6 microM and the dissociation constant for ARF1-guanosine 5',3-O-(thio)triphosphate (GTP[S]) was 7.4+/-2.2 microM. GAP2 was similar with a Km for ARF1-GTP of 5.4+/-1.2 microM and a dissociation constant for ARF1-GTP[S] of 4.8+/-0.3 micro...Continue Reading

Citations

Jan 20, 2005·The Journal of Cell Biology·Stella Y LeeVictor W Hsu
Nov 4, 2000·The Journal of Cell Biology·T R JacksonP A Randazzo
Jun 25, 2005·Molecular Biology of the Cell·Ya-Wen LiuFang-Jen S Lee
Jul 3, 1999·The Journal of Biological Chemistry·J Gaschet, V W Hsu
Jan 13, 2005·The Journal of Biological Chemistry·Nobutaka SaekiMitsuo Ikebe
May 23, 2013·Traffic·Frank AdolfFelix T Wieland
Feb 18, 1999·Molecular and Cellular Biology·J AndreevJ Schlessinger
Oct 16, 2002·The Journal of Cell Biology·Jia-Shu YangVictor W Hsu
Aug 17, 2011·The Journal of Biological Chemistry·Vincent PopoffFelix T Wieland
Sep 20, 2020·International Journal of Molecular Sciences·Fumio SakaneChiaki Murakami
Sep 24, 2014·Pharmacological Reviews·Ronald C BruntzH Alex Brown
May 14, 1999·Trends in Cell Biology·M G Roth
Jan 8, 2004·Cellular Signalling·Paul A Randazzo, Dianne S Hirsch

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