Lipid requirements for reconstitution of the delipidated beta-adrenergic receptor and the regulatory protein.
The role of lipids in the interaction of the beta-adrenergic receptor (R) with the regulatory protein (Gs) was investigated. Solubilized preparations of R and of Gs from turkey erythrocytes were delipidated by gel filtration. They were subsequently combined and reconstituted by the addition of various lipids. When reconstitution was carried out in the presence of soybean lipids, Gs could be fully activated via R by addition of hormone plus GTP gamma S. In contrast, purified phospholipids or a phospholipid fraction from soybean failed to produce an active system. Fractionation of soybean lipids revealed that acetone-soluble neutral lipids are essential for the reconstitution of a hormone responsive system. The acetone fraction could be replaced by specific neutral lipids such as alpha-tocopherol or cholesteryl arachidonate while a mixture of phosphatidylethanolamine, -choline and -serine satisfied the phospholipid requirement of the system.
Kirilovsky
J
J
Steiner-Mordoch
S
S
Selinger
Z
Z
Schramm
M
M
D000818:Animals; D004910:Erythrocyte Membrane / Q000032:analysis; D019204:GTP-Binding Proteins / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D007545:Isoproterenol / Q000494:pharmacology; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D015636:Magnesium Chloride; D008563:Membrane Lipids / Q000097:blood / Q000502:physiology*; D010743:Phospholipids / Q000502:physiology; D011943:Receptors, Adrenergic, beta / Q000502:physiology*; D013025:Glycine max; D013873:Thionucleotides / Q000494:pharmacology; D014422:Turkeys / Q000097:blood
D008563:Membrane Lipids; D010743:Phospholipids; D011943:Receptors, Adrenergic, beta; D013873:Thionucleotides; D015636:Magnesium Chloride; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; D008274:Magnesium; D007545:Isoproterenol