Molecular maps of red cell deformation: hidden elasticity and in situ connectivity.
Fluorescence-imaged micropipette aspiration was used to map redistribution of the proteins and lipids in highly extended human red blood cell membranes. Whereas the fluid bilayer distributed uniformly (+/- 10 percent), the underlying, solidlike cytoskeleton of spectrin, actin, and protein 4.1 exhibited a steep gradient in density along the aspirated projection, which was reversible on release from deformation. Quantitation of the cytoskeletal protein density gradients showed that skeletal elasticity is well represented by a grafted polymer network with a ratio of surface dilation modulus to shear modulus of approximately 2:1. Fractionally mobile integral proteins, such as band 3, and highly mobile receptors, such as CD59 as well as glycophorin C in protein 4.1-deficient cells, appeared to be squeezed out of areas dense in the underlying network and enriched in areas of network dilation. This complementary segregation demonstrates patterning of cell surface components by cytoskeletal dilation.
Joint Graduate Group in Bioengineering, University of California, Berkeley, San Francisco 94720.
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Discher
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Mohandas
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Evans
D000199:Actins / Q000097:blood; D001457:Anion Exchange Protein 1, Erythrocyte / Q000032:analysis; D001798:Blood Proteins / Q000032:analysis*; D003598:Cytoskeletal Proteins; D003599:Cytoskeleton / Q000737:chemistry / Q000502:physiology*; D004548:Elasticity; D004907:Erythrocyte Deformability; D004910:Erythrocyte Membrane / Q000737:chemistry* / Q000502:physiology; D004912:Erythrocytes / Q000737:chemistry / Q000166:cytology / Q000502:physiology*; D006021:Glycophorins / Q000032:analysis; D017261:Glycosylphosphatidylinositols / Q000097:blood; D006801:Humans; D008051:Lipid Bilayers; D008563:Membrane Lipids / Q000097:blood*; D008565:Membrane Proteins / Q000032:analysis; D008856:Microscopy, Fluorescence; D009479:Neuropeptides; D013049:Spectrin / Q000032:analysis
D000199:Actins; D001457:Anion Exchange Protein 1, Erythrocyte; D001798:Blood Proteins; D003598:Cytoskeletal Proteins; D006021:Glycophorins; D017261:Glycosylphosphatidylinositols; D008051:Lipid Bilayers; D008563:Membrane Lipids; D008565:Membrane Proteins; D009479:Neuropeptides; C028841:erythrocyte membrane band 4.1 protein; C120557:erythrocyte membrane protein band 4.1-like 1; D013049:Spectrin