Spectrin-protein 4.1-actin complex

The intricate interactions of the spectrin-protein 4.1-actin complex may be of central importance in maintaining the structural integrity of the red cell membrane. Two genetic disorders affecting the red cell membrane skeleton are hereditary spherocytosis and hereditary elliptocytosis. The former, the most common congenital form of hemolytic anemia in persons of northern European descent, exhibits an autosomal dominant inheritance pattern. The red blood cells are spherical, osmotically fragile, and considerably reduced in life span. They undergo... 

Figured. Diagrammatic representation of the red blood cell cytoskeletal-plasma membrane complex. Spectrin is made up of many homologous triple-helical segments joined by nonhelical regions (Speicher and Marchesi, 1984). Spectrin and actin require accessory proteins to form a membrane-associated network. (This diagram is constructed from data previously published for example, see Stryer, 1988 Davies and Lux, 1989 Bennett and Gilligan, 1993).

The spectrin cytoskeleton is connected to the membrane lipid bilayer by ankyrin, which interacts with (3-spectrin and the integral membrane protein, band 3. Band 4.2 helps to stabilize this connection. Band 4.1 anchors the spectrin skeleton with the membrane by binding the integral membrane protein glycophorin C and the actin complex, which has bound multiple spectrin dimers. 

Bundles of parallel actin filaments with uniform polarity. The microvilli of intestinal epithelial cells (enterocytes) are packed with actin filaments that are attached to the overlying plasma membrane through a complex composed of a 110-kD protein and calmodulin. The actin filaments are attached to each other through fimbrin (68 kD) and villin (95 kD). The actin bundles that emerge out of the roots of microvilli disperse horizontally to form a filamentous complex, the terminal web, in which several cytoskeletal proteins, spectrin (fodrin), myosin, actinin, and tropomyosin are present. Actin in the terminal web also forms a peripheral ring, which is associated with the plasma membrane on the lateral surfaces of the enterocyte (see Figure 5, p. 24). 

Protein 4.1, a globular protein, binds tightly to the tail end of spectrin, near the actin-binding site of the latter, and thus is part of a protein 4.1-spectrin-actin ternary complex. Protein 4.1 also binds to the integral proteins, glycophorins A and C, thereby attaching the ternary complex to the membrane. In addition, protein 4.1 may interact with certain membrane phospholipids, thus connecting the lipid bilayer to the cytoskeleton. 

Figure 3. Critical concentration behavior of actin self-assembly. For the top diagram depicting the macroscopic critical concentration curve, one determines the total amount of polymerized actin by methods that measure the sum of addition and release processes occurring at both ends. Examples of such methods are sedimentation, light scattering, fluorescence assays with pyrene-labeled actin, and viscosity measurements. Forthe bottom curves, the polymerization behavior is typically determined by fluorescence assays conducted under conditions where one of the ends is blocked by the presence of molecules such as gelsolin (a barbed-end capping protein) or spectrin-band 4.1 -actin (a complex prepared from erythrocyte membranes, such that only barbed-end growth occurs). Note further that the barbed end (or (+)-end) has a lower critical concentration than the pointed end (or (-)-end). This differential stabilization requires the occurrence of ATP hydrolysis to supply the free energy that drives subunit addition to the (+)-end at the expense of the subunit loss from the (-)-end.

A primary function of the SH3 domains is to form fimctional oligomeric complexes at defined subcellular sites, frequently in cooperation with other modular domains. SH3 domains are foimd in many proteins associated with the cytoskeleton or with the plasma membrane. Examples are the actin binding protein a-spectrin and myosin lb. Furthermore, SH3 interactions are involved in signal transduction in the Ras pathway (see Chapter 9). 

Fig. 4. Structure of striated muscle costameres and the DPC. A single membrane-associated costamere from a portion of a striated muscle fiber is magnified above to show the components of the dystrophin-associated protein complex that are involved in linking desmin intermediate filaments (IFs) to the muscle cell membrane. Additional actin-associated proteins present at these sites (including vinculin, talin, spectrin, and ankyrin) are not shown here. In addition to components of the DPC, plectin has also been localized to costameres, and likely contributes to linking desmin IFs to actin-associated structures.

A protein with a similar dumbell shape and structure is troponin C of skeletal muscles. Troponin C binds to a complex of proteins that assemble on the thin actin filaments of muscle fibers and control con-trachon in response to changes in the calcium ion con-centrahon (Chapter 19). Other proteins that contain EF-hand mohfs and are therefore responsive to Ca + include spectrin of cell membranes, clathrin light chains from coated vesicles, the extracellular osteonectin of bones and teeth, ° and a birch pollen anhgen. 2 Another group of 17 or more small SlOO EF-hand proteins play a variety of other roles. One of these, which has a high affinity for Zn +, has been named psoriasin because of its 5-fold or greater... 

Fig. 44.10. A generalized view of the erythrocyte cytoskeleton. A. The major protein, spectrin, is linked to the plasma membrane either through interactions with ankyrin and band 3, or with actin, band 4.1, and glycophorin. Other proteins in this complex, but not shown, are tropomyosin and adducin. B. A view from inside the cell, looking up at the cytoskeleton. This view displays the cross-linking of the sprectrin dimers to actin and band 3 anchor sites.

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