title: Composition of the ternary protein complex of the red cell membrane cytoskeleton.
abstract: The red cell membrane skeletal network is constructed from actin, spectrin and protein 4.1 in a molar ratio of actin subunits/spectrin heterodimer/protein 4.1 of 2:1:1. This represents saturation of the actin filaments, since incubation with extraneous spectrin and protein 4.1 leads to no binding of additional spectrin, either to the inner surface of ghost membranes or to lipid-free membrane cytoskeletons. Partial extraction of spectrin from the membrane is accompanied by release of actin under all conditions. Regardless of the proportion of spectrin extracted, the molar ratio of spectrin dimers/actin subunits is constant at 1:2. This is not the result of release or cooperative breakdown of whole lattice junctions from the network, for the number of actin filaments, judged by capacity to nucleate polymerisation of added G-actin, remains unchanged even when as much as 60% of the total spectrin has been lost. A similar 1:2:1 stoichiometry characterises the complex formed when G-actin is allowed to polymerise in the presence of varying amounts of spectrin and protein 4.1. When this complex is treated with the depolymerising agent, 1 M guanidine hydrochloride, it breaks down into smaller units of the same stoichiometry. After cross-linking these can be recovered from a gel-filtration column. Complexes prepared starting from G-actin appear to be much more stable than those formed when spectrin and protein 4.1 are bound to F-actin.
authors.affiliation: Medical Research Council Cell Biophysics Unit, King's College London, UK.
authors.forename: A
authors.initials: A
authors.lastname: Pekrun
authors.forename: J C
authors.initials: JC
authors.lastname: Pinder
authors.forename: S A
authors.initials: SA
authors.lastname: Morris
authors.forename: W B
authors.initials: WB
authors.lastname: Gratzer
mesh_terms: D000199:Actins / Q000032:analysis; D001704:Biopolymers / Q000032:analysis; D003598:Cytoskeletal Proteins / Q000032:analysis*; D003956:Dialysis; D004910:Erythrocyte Membrane / Q000032:analysis*; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis*; D011485:Protein Binding; D013049:Spectrin / Q000032:analysis; D013050:Spectrometry, Fluorescence
chemical_list: D000199:Actins; D001704:Biopolymers; D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D013049:Spectrin