Ras GTPase-activating protein physically associates with mitogenically active phospholipids.
The physical interaction between GTPase-activating protein (GAP) and lipids has been characterized by two separate analyses. First, bacterially synthesized GAP molecules were found to associate with detergent-mixed micelles containing arachidonic but not with those containing arachidic acid. This association was detected by a faster elution time during molecular exclusion chromatography. Second, GAP molecules within a crude cellular lysate were specifically retained by a column on which certain lipids had been immobilized. The lipids able to retain GAP on such columns were identical to those which were shown previously to be most active in blocking GAP activity. The association between lipids and GAP was dependent upon magnesium ions. Lipids unable to inhibit GAP activity were also unable to physically associate with GAP. The tight association of GAP with these lipids was predicted by and helps to rationalize their ability to inhibit GAP activity.
Tsai
M H
MH
Department of Molecular Biology, Cleveland Clinic Foundation, Ohio 44106.
Roudebush
M
M
Dobrowolski
S
S
Yu
C L
CL
Gibbs
J B
JB
Stacey
D W
DW
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003593:Cytoplasm / Q000378:metabolism; D004492:Edetic Acid / Q000494:pharmacology; D020690:GTPase-Activating Proteins; D006160:Guanosine Triphosphate / Q000378:metabolism; D007700:Kinetics; D008081:Liposomes; D051379:Mice; D008823:Micelles; D008970:Molecular Weight; D010743:Phospholipids / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism*; D013329:Structure-Activity Relationship; D020703:ras GTPase-Activating Proteins
D020690:GTPase-Activating Proteins; D008081:Liposomes; D008823:Micelles; D010743:Phospholipids; D011506:Proteins; D020703:ras GTPase-Activating Proteins; D006160:Guanosine Triphosphate; D004492:Edetic Acid