Rapid diffusion of spectrin bound to a lipid surface.
Human erythrocyte spectrin was labelled with the probe 5, 5'-disulfato-1-(6-hexanoic acid N-hydroxysuccinimide ester)-1'-ethyl-3,3,3',3'-tetramethylindocarbocyanine (Cy3). Cy3-spectrin was bound to the outer surface of dimyristoylphosphatidylcholine (DMPC) multilamellar vesicles and its diffusion measured by fluorescence recovery after photobleaching (FRAP). It was found that at 30 degrees C, above the lipid gel to liquid-crystalline phase transition of the lipids, Cy3-spectrin had an unexpectedly high diffusion coefficient D=(2.1+/-0.6)x10(-7)) cm2/s. At the phase transition, diffusion of Cy3-spectrin was only slightly lower; D=(1.3+/-0.3)x10(-7) cm2/s, whereas at 14 degrees C, well below the lipid phase transition, diffusion was found to be much slower with D=(3.1+/-0.12)x10(-9) cm2/s. The fast diffusion of Cy3-spectrin on the lipid surface implies that the individual bonds which bind spectrin to the lipid surface must rapidly be made and broken. In the light of these results, spectrin-lipid interactions alone appear unlikely to have any significant role in supporting the cell membrane. Probably, the interactions serve only to localise the spectrin at the inner lipid surface in order to facilitate formation of the cytoskeleton.
O'Toole
P J
PJ
Department of Biological Sciences, University of Essex, Wivenhoe Park, Colchester, Essex CO4 3SQ, UK. pjotoo@essex.ac.uk
Wolfe
C
C
Ladha
S
S
Cherry
R J
RJ
D002232:Carbocyanines; D004058:Diffusion; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D005456:Fluorescent Dyes; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry; D013049:Spectrin / Q000737:chemistry*; D013499:Surface Properties; D013696:Temperature
D002232:Carbocyanines; D005456:Fluorescent Dyes; D008055:Lipids; D008081:Liposomes; C089187:cyanine dye 3; D013049:Spectrin; D004134:Dimyristoylphosphatidylcholine