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