Membrane interaction of erythroid spectrin: surface-density-dependent high-affinity binding to phosphatidylethanolamine.
Density-dependent spectrin binding to dimyristoylphosphatidylcholine/dimyristoylphosphatidylethanolamine (DMPC/DMPE) small uni-lamellar vesicles (SUVs) has been directly evaluated in this work from the increase in the extent of quenching of the tryptophan fluorescence of spectrin at two different temperatures, above and below the main phase transition temperatures (Tm). Results from the binding studies of spectrin to phospholipid SUVs indicated that the binding dissociation constant Kd, increased from 45 +/- 7 nM in pure DMPC SUVs to 219 +/- 20 nM in DMPC/DMPE (50:50) SUVs, both in the gel and liquid crystalline phase. However, in pure DMPE SUVs the Kd decreased drastically to 0.7 +/- 0.2 nM in the gel phase at 18 degrees C and to 2.6 +/- 0.7 nM in the fluid phase at 55 degrees C indicating a high affinity binding of spectrin for the bilayer-forming DMPE. The maximum extent of phospholipid-induced quenching and the number of spectrin molecules associated with one SUV particle, evaluated in the present work, led to a model in DMPC/DMPE bilayer membranes indicating the PE-binding site of spectrin to localize at one of the terminal domains of the dimeric spectrin. A direct evidence of the localization of the PE-binding site at one of the terminal ends of the spectrin dimer also came from electron microscopic observation in fluid membranes made of bovine brain PE.
Ray
Sibnath
S
Biophysics Division, Saha Institute of Nuclear Physics, Kolkata, India.
Chakrabarti
Abhijit
A
D000818:Animals; D002417:Cattle; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry / Q000378:metabolism*; D004912:Erythrocytes / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D010714:Phosphatidylethanolamines / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D012756:Sheep; D013049:Spectrin / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013499:Surface Properties; D013696:Temperature; D013816:Thermodynamics; D014364:Tryptophan / Q000737:chemistry
D008051:Lipid Bilayers; D010714:Phosphatidylethanolamines; D013049:Spectrin; D014364:Tryptophan; D004134:Dimyristoylphosphatidylcholine; C018525:1,2-dimyristoylphosphatidylethanolamine