Resolved phospholipase D activity is modulated by cytosolic factors other than Arf.

作者: William D. Singer , H. Alex Brown , Gary M. Bokoch , Paul C. Sternweis

DOI: 10.1074/JBC.270.25.14944

关键词:

摘要: Phospholipase D, which has been extracted from porcine brain membranes and chromatographically enriched 100-fold, was activated better by impure preparations of Arf than purified or recombinant Arf. Examination cytosol with this preparation PLD activity revealed at least three stimulatory components. One these is the first cytoplasmic factor. A second peak PLD-stimulating (cytoplasmic factor II, CFII) resolved anion exchange gel filtration. This CFII can be further separated into multiple activities chromatography heparin-agarose. The were differentiated their properties as measured in absence presence guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) alone added GTP S. While all pools stimulated to some degree showed synergistic activation when administered conjunction Arf, they could classified two groups distinct behavior. When used together, respective PLD. set contained RhoA monomeric G protein. Recombinant show that it indeed activate act synergistically proteins. related protein, Cdc42, also effective. devoid and, contrast group, demonstrated significant stimulating guanine nucleotides. These data indicate modulated several cytosolic factors Arf-sensitive may represent a complex regulated an interactive fashion variety cellular signaling events.

参考文章(34)
M M Billah, J C Anthes, The regulation and cellular functions of phosphatidylcholine hydrolysis. Biochemical Journal. ,vol. 269, pp. 281- 291 ,(1990) , 10.1042/BJ2690281
Xuemin Wang, Liwen Xu, Ling Zheng, Cloning and expression of phosphatidylcholine-hydrolyzing phospholipase D from Ricinus communis L. Journal of Biological Chemistry. ,vol. 269, pp. 20312- 20317 ,(1994) , 10.1016/S0021-9258(17)31993-2
Paul A. Randazzo, Ofra Weiss, Richard A. Kahn, Preparation of recombinant ADP-ribosylation factor. Methods in Enzymology. ,vol. 219, pp. 362- 369 ,(1992) , 10.1016/0076-6879(92)19036-6
K C Malcolm, A H Ross, R G Qiu, M Symons, J H Exton, Activation of rat liver phospholipase D by the small GTP-binding protein RhoA. Journal of Biological Chemistry. ,vol. 269, pp. 25951- 25954 ,(1994) , 10.1016/S0021-9258(18)47140-2
J H Exton, Signaling through phosphatidylcholine breakdown. Journal of Biological Chemistry. ,vol. 265, pp. 1- 4 ,(1990) , 10.1016/S0021-9258(19)40184-1
T H Chuang, B P Bohl, G M Bokoch, Biologically active lipids are regulators of Rac.GDI complexation. Journal of Biological Chemistry. ,vol. 268, pp. 26206- 26211 ,(1993) , 10.1016/S0021-9258(19)74301-4
J K Northup, M D Smigel, A G Gilman, The guanine nucleotide activating site of the regulatory component of adenylate cyclase. Identification by ligand binding. Journal of Biological Chemistry. ,vol. 257, pp. 11416- 11423 ,(1982) , 10.1016/S0021-9258(18)33775-X
E.P. Bowman, D.J. Uhlinger, J.D. Lambeth, Neutrophil phospholipase D is activated by a membrane-associated Rho family small molecular weight GTP-binding protein. Journal of Biological Chemistry. ,vol. 268, pp. 21509- 21512 ,(1993) , 10.1016/S0021-9258(20)80570-5
P C Sternweis, J D Robishaw, Isolation of two proteins with high affinity for guanine nucleotides from membranes of bovine brain Journal of Biological Chemistry. ,vol. 259, pp. 13806- 13813 ,(1984) , 10.1016/S0021-9258(18)89817-9
H. Alex Brown, Paul C. Sternweis, Stimulation of phospholipase D by ADP-ribosylation factor. Methods in Enzymology. ,vol. 257, pp. 313- 324 ,(1995) , 10.1016/S0076-6879(95)57035-7