Striated paracrystals that contain HMWP, the homolog of actin-binding protein and filamin from HeLa cells.
HMWP (high molecular weight protein), a high molecular weight actin binding protein, was previously isolated from HeLa cells; its physical properties, amino acid composition, and intracellular localization indicated its homology with actin-binding protein and filamin [Weihing, 1982, 1983]. We now report the identification of HMWP in striated paracrystals. Purified HMWP is incubated at 25 degrees C and subjected to negative staining with uranyl acetate. Examination by electron microscopy reveals long, striated paracrystals formed from filaments a few nanometers in diameter that lie parallel to the long axis of the paracrystal. At intervals of about 200 nm, the filaments are crossed by granular aggregates, accounting for the striated appearance. Treatment of the paracrystals with an affinity-purified antibody to HMWP decorates the filaments; such decorations are not observed if nonimmune goat IgG or phosphate-buffered saline are substituted for the antibody. Electron microscopic and electrophoretic analysis of paracrystals sedimented onto grids by centrifugation at 864 g reveals that the grids are covered with paracrystals and the major polypeptide present on grids centrifuged in parallel is HMWP. Taken together, these data indicate that the filaments of the paracrystals contain elongated molecules of HMWP. Additional experiments are needed to decide if the paracrystals form by self-association between HMWP molecules or by association with one or more of the minor polypeptides that remain in the purified HMWP.
D002352:Carrier Proteins / Q000302:isolation & purification*; D003285:Contractile Proteins / Q000302:isolation & purification*; D064448:Filamins; D018260:Gelsolin; D006367:HeLa Cells / Q000032:analysis; D006801:Humans; D008840:Microfilament Proteins; D008854:Microscopy, Electron; D008970:Molecular Weight
D002352:Carrier Proteins; D003285:Contractile Proteins; D064448:Filamins; D018260:Gelsolin; D008840:Microfilament Proteins; C016953:brevin