Surface localization of the sea urchin egg receptor for sperm

The sea urchin egg receptor for sperm is thought to be involved in species-specific sperm-egg interactions at the egg surface. Recent revisions in the deduced amino acid sequence of the cloned cDNAs indicate that the protein encoded does not possess the common structural hallmarks of a membrane protein. Thus, investigation of the localization and association of the protein with the egg surface is crucial. We describe and characterize a new monoclonal antibody raised against recombinant sperm receptor protein. This antibody, in conjunction with several polyclonal antibodies, was used to study the receptor protein in eggs. Immunoprecipitation studies indicated that the antibodies recognize the high Mr (ca. 350 K) sperm receptor protein which copurified with egg plasma membrane-vitelline layer complexes. The sperm receptor protein was solubilized only by detergents and not by treatments designed to solubilize peripherally associated or lipid-anchored membrane proteins, suggesting a tight association with the membrane fraction. Confocal immunofluorescence microscopy of live eggs indicated surface staining. Finally, lysylendoproteinase C treatment of live eggs resulted in a loss of the high Mr receptor protein epitopes, and the concomitant release of a 70-kDa proteolytic fragment, which correlated with a reduced ability of the eggs to be fertilized. Taken together, these data indicate that at least some fraction of the sperm receptor protein is present on the egg surface, a requisite locale for a sperm binding protein.     

Cytoplasmic tail deletion of T cell receptor (TCR) beta-chain results in its surface expression as glycosylphosphatidylinositol-anchored polypeptide on mature T cells in the absence of TCR-alpha

Surface expression of the T cell antigen receptor (TCR) in mature T cells requires the association of a variable heterodimer (alpha.beta or gamma.delta) with six invariant CD3 polypeptides (gamma, delta, epsilon-epsilon, zeta-zeta, or zeta-eta). We described here that deletion of the cytoplasmic tail polypeptide sequence (Lys-Lys-Lys-Asn-Ser) of TCR beta-chain (beta CT) results in expression of the truncated beta-chain on the surface of a mature T cell hybridoma line, in the absence of TCR-alpha, as a glycophosphatidylinositol (GPI)-anchored monomeric polypeptide. The GPI-anchored TCR-beta CT is not associated with CD3-epsilon and is incapable of conventional signal transduction. Association with TCR-alpha prevents beta CT from GPI-linkage formation. The alpha beta CT heterodimer binds the CD3 polypeptides, and the resultant TCR alpha beta CT/CD3 complex is capable of signal transduction. Our data show that a signal sequence for GPI-linkage formation is present in TCR-beta, and this alternative membrane anchoring mechanism can be utilized by beta-chain polypeptide lacking the CT sequence. We conclude therefore that in the absence of TCR-alpha expression, the beta-chain CT sequence plays an essential function in hindering GPI-linkage formation, thereby preventing escape of incompletely assembled TCR beta-chain to the cell surface of mature T cells.     

The stationary-phase-exit defect of cydC (surB) mutants is due to the lack of a functional terminal cytochrome oxidase

The surB gene was identified as a gene product required for Escherichia coli cells to exit stationary phase at 37 degrees C under aerobic conditions. surB was shown to be the same as cydC, whose product is required for the proper assembly and activity of cytochrome d oxidase. Cytochrome d oxidase, encoded by the cydAB operon, is one of two alternate terminal cytochrome oxidases that function during aerobic electron transport in E. coli. Mutations inactivating the cydAB operon also cause a temperature-sensitive defect in exiting stationary phase, but the phenotype is not as severe as it is for surB mutants. In this study, we examined the phenotypes of surB1 delta(cydAB) double mutants and the ability of overexpression of cytochrome o oxidase to suppress the temperature-sensitive stationary-phase-exit defect of surB1 and delta(cydAB) mutants and analyzed spontaneous suppressors of surB1. Our results indicate that the severe temperature-sensitive defect in exiting stationary phase of surB1 mutants is due both to the absence of terminal cytochrome oxidase activity and to the presence of a defective cytochrome d oxidase. Membrane vesicles prepared from wild-type, surB1, and delta(cydAB) strains produced superoxide radicals at the same rate in vitro. Therefore, the aerobic growth defects of the surB1 and delta(cydAB) strains are not due to enhanced superoxide production resulting from the block in aerobic electron transport.     

Controlled oxygen incorporation in indium gallium arsenide and indium phosphide grown by metalorganic vapor phase epitaxy

The defect engineering in metalorganic vapor phase epitaxy In_xGa_1-xAs and InP by controlled oxygen doping using diethyl aluminum ethoxide (DEALO) was developed in this study. DEALO doping has led to the incorporation of Al and O, and the compensation of shallow Si donors in In_xGa_1−xAs: Si with 0 ≤ x ≤ 0.25. With the same DEALO mole fraction during growth, the incorporation of Al and O was found to be independent of x, but the compensation of Si donors decreases with increasing In content. Deep level transient spectroscopy analysis on a series of In_xGa_1-xAs: Si. samples with 0 ≤ x ≤ 0.18 revealed that oxygen incorporation led to a set of deep levels, similar to those found in DEALO doped GaAs. As the In composition was increased, one or more of these deep levels became resonant with the conduction band and led to a high electron concentration in oxygen doped In_0.53Ga_0.47As. Low temperature photoluminescence emission measurements at 12K on the same set of samples revealed the quenching of the near-band edge peak, and the appearance of new oxygen-induced emission features. DEALO doping in InP has also led to the incorporation of Al and O, and the compensation of Si donors due to oxygen-induced multiple deep levels.     

Adolescents in stepfamilies: Developmental family interventions.

Discusses issues and interventions for working with adolescents who live in stepfamilies. A developmental perspective, using psychoeducation and brief strategic intervention approaches, is proposed for working with stepfamilies. Six major issues for adolescents in stepfamilies are discussed: developmental issues, sexuality issues, parent–child relationships, parenting in stepfamilies, nonresidential parent–child issues, and changes in visitation and custody. Case illustrations and suggested interventions are presented for each of these areas. (PsycINFO Database Record (c) 2010 APA, all rights reserved)