Glycosylation of nucleocytoplasmic proteins: signal transduction and O-GlcNAc

The dynamic glycosylation of serine or threonine residues on nuclear and cytosolic proteins by O-linked beta-N-acetylglucosamine (O-GlcNAc) is abundant in all multicellular eukaryotes. On several proteins, O-GlcNAc and O-phosphate alternatively occupy the same or adjacent sites, leading to the hypothesis that one function of this saccharide is to transiently block phosphorylation. The diversity of proteins modified by O-GlcNAc implies its importance in many basic cellular and disease processes. Here we systematically examine the current data implicating O-GlcNAc as a regulatory modification important to signal transduction cascades.     

Dimerization of the extracellular domain of the human growth hormone receptor by a single hormone molecule

Human growth hormone (hGH) forms a 1:2 complex with the extracellular domain of its receptor-binding protein (hGHbp) as studied by crystallization, size exclusion chromatography, calorimetry, and a previously undescribed fluorescence quenching assay. These and other experiments with protein engineered variants of hGH have led to the identification of the binding determinants for two distinct but adjacent sites on hGH for the hGHbp, and the data indicated that there are two overlapping binding sites on the hGHbp for hGH. Furthermore, the binding of hGH to the hGHbp occurred sequentially; a first hGHbp molecule bound to site 1 on hGH and then a second hGHbp bound to site 2. Hormone-induced receptor dimerization is proposed to be relevant to the signal transduction mechanism for the hGH receptor and other related cytokine receptors.     

Development of an analytical scheme for simazine and 2,4-D in soil and water runoff from ornamental plant nursery plots

The transport and fate of pesticides applied to ornamental plant nursery crops are not well documented. Methodology for analysis of soil and water runoff samples concomitantly containing the herbicides simazine (1-chloro-4,6-bis(ethylamino)-s-triazine) and 2,4-D ((2,4-dichlorophenoxy)acetic acid) was developed in this research to investigate the potential for runoff and leaching from ornamental nursery plots. Solid-phase extraction was used prior to analysis by gas chromatography and liquid chromatography. Chromatographic results were compared with determination by enzyme-linked immunoassay analysis. The significant analytical contributions of this research include (1) the development of a scheme using chromatographic mode sequencing for the fractionation of simazine and 2,4-D, (2) optimization of the homogeneous derivatization of 2,4-D using the methylating agent boron trifluoride in methanol as an alternative to in situ generation of diazomethane, and (3) the practical application of these techniques to field samples.     

Plant and animal species composition and heavy metal content in fly ash ecosystems

Plant and animal species present on a coal fly ash slurry pond site and a dry deposit site were surveyed and sampled during a two-day period in October. Elemental analyses were determined for most of the species encountered. A total of 48 plant species were observed on the two sites, with 35 species on the wet site, and 20 on the dry site. Eighteen terrestrial and 7 aquatic animal species were found on the wet site, exclusive of vertebrates which were not studied with the exception of a carp (Cyprinus carpio). Eleven terrestrial invertebrates and one aquatic species were observed on the dry site. Neutron activation analysis was carried out for: Se, Hg, Cr, Ni, Zn, Co, Sb, Cd, and As. Using literature values for phytotoxicity, we conclude that, in general, plants did not accumulate toxic levels of metals. Only one plant (Impatiens biflora Willd.) showed a significant level of Cd. Of 20 plants analyzed on the wet site, 10 had excessive Se concentrations (>5 ppm); on the dry site 6 out of 18 had high Se values. In animals (Gryllus sp.; Melanoplus sp.; Trachelipus sp.; Lumbricus terrestris; Physa integra; Cyprinus carpio) the trace metal concentration was generally in between that of control animals and that of the fly ash itself. One exception included Zn, which, although the most variable element examined, was concentrated in all the terrestrial animals to levels higher than in fly ash. Crickets are the most consistent bioconcentrators with Cr, Se, and Zn at higher levels than for control animals. All animal species studied accumulated Se compared to controls.