title: Molecular analysis of insertion/deletion mutations in protein 4.1 in elliptocytosis. I. Biochemical identification of rearrangements in the spectrin/actin binding domain and functional characterizations.
abstract: Protein 4.1 (80 kD) interacts with spectrin and short actin filaments to form the erythrocyte membrane skeleton. Mutations of spectrin and protein 4.1 are associated with elliptocytosis or spherocytosis and anemia of varying severity. We analyzed two mutant protein 4.1 molecules associated with elliptocytosis: a high molecular weight 4.1 (95 kD) associated with mild elliptocytosis without anemia, and a low molecular weight 4.1 (two species at 68 and 65 kD) associated with moderate elliptocytosis and anemia. 4.1(95) was found to contain a approximately 15-kD insertion adjacent to the spectrin/actin binding domain comprised, at least in part, of repeated sequence. 4.1(68/65) was found to lack the entire spectrin-actin binding domain. The mechanical stability of erythrocyte membranes containing 4.1(95) was identical to that of normal membranes, consistent with the presence of an intact spectrin-actin binding domain in protein 4.1. In contrast, membranes containing 4.1(68/65) have markedly reduced mechanical stability as a result of deleting the spectrin-actin binding domain. The mechanical stability of these membranes was improved following reconstitution with normal 4.1. These studies have thus enabled us to establish the importance of the spectrin-actin binding domain in regulating the mechanical stability of the erythrocyte membrane.
authors.affiliation: Department of Laboratory Medicine, Yale University School of Medicine, New Haven, Connecticut 06510.
authors.forename: S L
authors.initials: SL
authors.lastname: Marchesi
authors.forename: J
authors.initials: J
authors.lastname: Conboy
authors.forename: P
authors.initials: P
authors.lastname: Agre
authors.forename: J T
authors.initials: JT
authors.lastname: Letsinger
authors.forename: V T
authors.initials: VT
authors.lastname: Marchesi
authors.forename: D W
authors.initials: DW
authors.lastname: Speicher
authors.forename: N
authors.initials: N
authors.lastname: Mohandas
mesh_terms: D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D001665:Binding Sites; D003598:Cytoskeletal Proteins; D015180:Electrophoresis, Gel, Two-Dimensional; D004612:Elliptocytosis, Hereditary / Q000235:genetics*; D004907:Erythrocyte Deformability; D004910:Erythrocyte Membrane / Q000502:physiology; D004912:Erythrocytes / Q000032:analysis; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D009479:Neuropeptides; D010446:Peptide Fragments / Q000032:analysis; D013049:Spectrin / Q000378:metabolism
chemical_list: D000199:Actins; D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D009479:Neuropeptides; D010446:Peptide Fragments; C028841:erythrocyte membrane band 4.1 protein; C120557:erythrocyte membrane protein band 4.1-like 1; D013049:Spectrin