title: Moesin, ezrin, and p205 are actin-binding proteins associated with neutrophil plasma membranes.
abstract: Actin-binding proteins in bovine neutrophil plasma membranes were identified using blot overlays with 125I-labeled F-actin. Along with surface-biotinylated proteins, membranes were enriched in major actin-binding polypeptides of 78, 81, and 205 kDa. Binding was specific for F-actin because G-actin did not bind. Further, unlabeled F-actin blocked the binding of 125I-labeled F-actin whereas other acidic biopolymers were relatively ineffective. Binding also was specifically inhibited by myosin subfragment 1, but not by CapZ or plasma gelsolin, suggesting that the membrane proteins, like myosin, bind along the sides of the actin filaments. The 78- and 81-kDa polypeptides were identified as moesin and ezrin, respectively, by co-migration on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoprecipitation with antibodies specific for moesin and ezrin. Although not present in detectable amounts in bovine neutrophils, radixin (a third and closely related member of this gene family) also bound 125I-labeled F-actin on blot overlays. Experiments with full-length and truncated bacterial fusion proteins localized the actin-binding site in moesin to the extreme carboxy terminus, a highly conserved sequence. Immunofluorescence micrographs of permeabilized cells and cell "footprints" showed moesin co-localization with actin at the cytoplasmic surface of the plasma membrane, consistent with a role as a membrane-actin-linking protein.
authors.affiliation: Worcester Foundation for Experimental Biology, Shrewsbury, Massachusetts 01545, USA.
authors.forename: K
authors.initials: K
authors.lastname: Pestonjamasp
authors.forename: M R
authors.initials: MR
authors.lastname: Amieva
authors.forename: C P
authors.initials: CP
authors.lastname: Strassel
authors.forename: W M
authors.initials: WM
authors.lastname: Nauseef
authors.forename: H
authors.initials: H
authors.lastname: Furthmayr
authors.forename: E J
authors.initials: EJ
authors.lastname: Luna
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001798:Blood Proteins / Q000378:metabolism; D002417:Cattle; D003598:Cytoskeletal Proteins; D003599:Cytoskeleton / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000378:metabolism* / Q000648:ultrastructure; D010750:Phosphoproteins / Q000378:metabolism*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D017384:Sequence Deletion; D017385:Sequence Homology; D000099149:Ezrin
chemical_list: D001798:Blood Proteins; D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D008840:Microfilament Proteins; D010750:Phosphoproteins; D011506:Proteins; D000099149:Ezrin; C071058:moesin; C071776:radixin