Fast Atom Bombardment-High Field Magnet Mass Spectrometry of 6000 Dalton polypeptides

Using a combination of Fast Atom Bombardment lonisation and High Field Magnet — Mass Spectrometry we demonstrate here the ability to observe, for the first time, the accurate molecular weights of polypeptides or other important bioorganic substances up to 6000 Dalton in size. This claim is substantiated with data on intact unreduced Insulin and on the intact sweet protein Monellin, which consists of three polypeptide chains.     

Resistance to moloney murine sarcoma virus-induced tumorigenesis in NK-deficient beige mice

C57BL/6J-bg^J$\text{bg}\text{bg}$ mice are reported to be less susceptible to tumor induction by threshold doses of Moloney murine sarcoma virus than their +/bg littermates, and there are no significant differences between $\text{bg}\text{bg}$ and +/bg mice in which tumors were induced with respect to tumor latency, size, and regression rate. The difference in tumor frequency cannot be accounted for by M-MSV boosting of activity in $\text{bg}\text{bg}$ mice or by depression of activity in +/bg animals.     

Biogeographic comparisons of marine algal polyphenolics: evidence against a latitudinal trend

Summary Marine allelochemicals generally are present in greater quantity and diversity in tropical than in temperate regions. Marine algal polyphenolics have been reported as an apparent exception to this biogeographic trend, with literature values for phenolic concentrations significantly higher in temperate than in tropical brown algae. In contrast, our results, the first reported for Caribbean brown algae (orders Dictyotales and Fucales), show that many species have high phenolic levels. In addition, both our study and previous studies with north temperate and tropical species demonstrate that there is marked variation in algal phenolic levels within species from different locations. We conclude that high phenolic concentrations occur in species from both temperate and tropical regions, indicating that latitude alone is not a reasonable predictor of plant phenolic concentrations.     

Immunocytolocalization of glutamine synthetase in mesophyll and phloem of leaves ofSolanum tuberosum L.

Summary Localization of glutamine synthetase inSolanum tuberosum leaves was investigated by techniques of Western tissue printing and immunogold electron microscopy. Anti-GS antibodies used in immunolocalization recognize two peptides (45 kDa and 42 kDa) on Western blots. Antibody stained tissue prints on nitrocellulose membranes allowed low resolution localization of GS. Immunostaining was most evident in the adaxial phloem of the leaf midribs and petiole veins. High-resolution localization of glutamine synthetase by immunogold electron microscopy revealed that this enzyme occurs in both the chloroplasts and the cytosol ofS. tuberosum leaf cells. However, GS was specifically associated with the chloroplasts of mesophyll cells and with the cytoplasm of phloem companion cells. The evidence for cell-specific localization of chloroplast and cytosolic GS presented here agrees with the recently reported cell-specific pattern of expression of GUS reporter gene, directed by promoters for chloroplast and cytosolic GS form in tobacco transgenic plants. These data provide additional clues to the interpretation of the functional role of these different isoenzymes and its relationship with their specific localization.Abbreviations BSA bovine serum albumin - EM electron microscope - GOGAT glutamate synthase - GS glutamine synthetase - GUS -glucuronidase - IgG immunoglobulin - PBS phosphate buffer saline - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis     

Contrasts between subsurface microbial communities and their metabolic adaptation to polycyclic aromatic hydrocarbons at a forested and an urban coal-tar disposal site

The abundance and distribution of microorganisms and their potential for mineralizing polycyclic aromatic hydrocarbons (PAHs) were measured in subsurface sediment samples at two geographically separate buried coal-tar sites. At a relatively undisturbed forested site in the northeastern United States, metabolic adaptation to the PAHs was evident: Radiolabeled naphthalene and phenanthrene were converted to ¹⁴CO₂ in core material from inside but not outside a plume of groundwater contamination. However, at the urban site in the midwestern United States these PAHs were mineralized in sediments from both contaminated and uncontaminated boreholes. Thus, clear qualitative evidence showing an adaptational response by the subsurface microbial community was not obtained at the urban site. Instead, subtler clues suggesting metabolic adaptation by subsurface microorganisms from the urban site were discerned by comparing lag periods and extents of ¹⁴CO₂ production from radiolabeled PAHs added to samples from contaminated and uncontaminated boreholes. Despite slightly higher PAH mineralization activity in contaminated borehole samples, p-hydroxybenzoate was mineralized equally in all samples from the urban site regardless of location. No striking trends in the abundances of actinomycetes, fungi, and either viable or total bacteria were encountered. However, colonies of the soil bacterium, Bacillus mycoides, were detected on enumeration plates of several samples from unsaturated and saturated zones in both urban boreholes. Furthermore, other common soil bacteria, Myxococcus xanthus and Chromobacterium violaceum, were identified in samples from the uncontaminated urban borehole. The occurrence of bacteria usually restricted to surface soil, combined with the observation of fragments of building materials in many of the core samples, suggested that past excavation and backfilling operations may have caused mixing of surface soil with subsurface materials at the urban site. We speculate that this mixing, as well as non-coal-tar-derived sources of PAHs, contributed to the PAH-mineralizing activity present in the sediment samples from the uncontaminated urban borehole.     

Microbial biomass and activity in subsurface sediments from Vejen,Denmark

Subsurface sediment samples were collected from 4 to 31 m below landsurface in glacio-fluvial sediments from the Quaternary period. The samples were described in terms of pH, electrical conductivity, chloride concentration, organic matter content, and grain size distribution. Viable counts of bacteria varied from 0.5 to 1,203 x 103 colony forming units/g dry weight (gdw); total numbers of bacteria acridine orange direct counts (AODC) varied from 1.7 to 147 × 10⁷ cells/gdw; growth rates (incorporation of [³H]-thymidine) varied from 1.4 to 60.7 × 10⁴ cells/(gdw · day); and rate constants for mineralization of ¹⁴C-labelled compounds varied from 0.2 to 2.3 × 10^–3 ml/(dpm · day) for acetate, and from 0 to 2.0 × 10^–3 ml/(dpm · day) for phenol. Sediment texture influenced the total number of bacteria and potential for mineralization; with increasing content of clay and silt and decreasing content of sand, AODC increased and the mineralization rate declined. Intrinsic permeability calculated from grain size correlated positively with mineralization rate for acetate. Statistical correlation analysis showed high correlations between some of the abiotic parameters, but it was not possible to point out a single abiotic parameter that could explain the variation of size and activity of the microbial population. The microbial data obtained in these geologically young sediments were compared to literature data from older sediments, and this comparison showed that age and type of geological formation might be important for the size and activity of the microbial populations.Offprint requests to: H.-J. Albrechtsen.     

Phenylalanine 4-monooxygenase from bovine and rat liver: Some physical and chemical properties

Phenylalanine 4-monooxygenase was purified from bovine liver using a modification of the procedure developed for the rat liver enzyme (Shiman, R., Gray, D. W., and Pater, A. 1979. J. Biol. Chem. 254:11300–11306). The enzyme preparation appeared essentially homogeneous on polyacrylamide gel electrophoresis under non-denaturing conditions. Electrophoresis in the presence of dodecyl sulfate revealed that about 95% of the protein had a mobility, corresponding to M_r=51,000. The remaining 5% was recovered in two minor bands corresponding to M_r of about 35,000 and 15,000 and is likely to result from limited proteolysis of the native enzyme with dissociation of the fragments on denaturation by detergent. The enzyme comigrated with the rat liver enzyme on polyacrylamide gel electrophoresis in both systems studied. No significant difference was observed between the amino acid composition of the bovine and rat liver enzyme, in the reactivity of their sulfhydryl groups or in their iron content (i.e. 1.5–3.0 iron atoms per peptide chain of M_r=50,000). Both enzymes contained less than 0.01 copper atom per peptide chain. The enzymes were inhibited in a similar manner by the chelator bathophenanthroline disulfonate (selective for iron and copper), but not by bathocuproine disulfonate (specific for copper). The results indicate that the bovine and rat liver enzymes are closely similar and that iron, but not copper, is essential for enzyme activity. High performance size-exclusion liquid chromatography revealed that both native enzymes exist in different oligomeric forms, but further studies are required to understand the physicochemical basis for this phenomenon.Abbreviations Bathophenanthroline 4,7-diphenyl-1, 10-phenanthroline - bathocuproine 2,9-dimethyl-4,7-diphenyl-1, 10-phenanthroline - Hepes N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - dansyl 1-dimethylaminonaphthalene-5-sulfonyl - DMPH₄ 2-amino-4-hydroxy-6,7-dimethyl-5,6,7,8-tetrahydropteridine - M_r relative molecular mass     

INCREASE IN SIZE AND CELL NUMBER OF LATERAL ROOT PRIMORDIA IN THE PRIMARY OF INTACT PLANTS AND IN EXCISED ROOTS OF PISUM SATIVUM AND VICIA FABA

Increase in cell number, and in anlage volume and length have been investigated during the development of lateral root primordia in roots of intact plants of Pisum sativum and Vicia faba and in excised roots of both species cultured in White's medium supplemented with 2% sucrose. With the exception of primordia in excised roots of Vicia, the general equation which best described increase in each aspect of primoridium growth measured against time was that for exponential growth. When the times necessary for cell number and primordium volume and length to double were determined at intervals over the period of development studied, however, they were found to vary. Similarly, estimates of the size of the proliferative fraction of cells at different times during anlage development indicated that this index of meristematic activity also fluctuated over the developmental period investigated, i.e., increase in cell number and in primordium volume and length do not occur in a truly exponential fashion as the primordia increase in size and cell number. One difference between anlage development in the roots of intact plants and in those grown in culture was that whereas the former primordia completed their development and emerged as lateral roots over the period of the investigation, the latter did not. Moreover, cell doubling time and anlage volume and length doubling times were longer, and the proliferation fraction of cells lower, over the whole period of, and at intervals during, primordium development in the excised roots compared with the results obtained for the roots of the corresponding intact plants.