A photoactivable phospholipid analogue that specifically labels membrane cytoskeletal proteins of intact erythrocytes.
A radioactive photoactivable analogue of phosphatidylethanolamine, 2-(2-azido-4-nitro-benzoyl)-1-acyl-sn-glycero-3-phospho[14C]ethanolamine ([14C]AzPE), was synthesized. Upon incubation with erythrocytes in the dark, about 90% of [14C]AzPE spontaneously incorporated into the cells; of this fraction, about 90% associated with the membrane, all of it noncovalently. Upon photoactivation, 3-4% of the membrane-associated probe was incorporated into protein. Analysis of this fraction by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, as well as extraction of labeled membranes with alkali or detergent, showed that the probe preferentially labeled cytoskeletal proteins. [14C]AzPE appears to be a useful tool for the study of lipid-protein interactions at the cytoplasmic face of the plasma membrane of intact cells.
Pradhan
D
D
Department of Molecular and Cell Biology, Pennsylvania State University 16802.
Williamson
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P
Schlegel
R A
RA
D000345:Affinity Labels / Q000378:metabolism*; D002250:Carbon Radioisotopes; D003598:Cytoskeletal Proteins / Q000097:blood* / Q000302:isolation & purification; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006801:Humans; D007202:Indicators and Reagents; D007700:Kinetics; D008565:Membrane Proteins / Q000097:blood* / Q000302:isolation & purification; D010714:Phosphatidylethanolamines / Q000097:blood* / Q000138:chemical synthesis; D010782:Photolysis
D000345:Affinity Labels; D002250:Carbon Radioisotopes; D003598:Cytoskeletal Proteins; D007202:Indicators and Reagents; D008565:Membrane Proteins; D010714:Phosphatidylethanolamines; C061749:2-(2-azido-4-nitrobenzoyl)-1-myristoylglycero-3-phosphoethanolamine