Phosphorylation of liver pyruvate kinase by Ca++/calmodulin-dependent protein kinase: characterization of two phosphorylation sites

Rat liver pyruvate kinase is phosphorylated by calcium/calmodulin-dependent protein kinase II at serine and threonine residues in a 3-4 kDa CNBr fragment located near the amino terminus. The two sites of phosphorylation were separated by reverse-phase HPLC of a thermolysin digest. Sequence analysis established the sites of phosphorylation as follows: Leu-Arg-Arg-Ala-Ser(PO4)-Val-Ala-Gln-Leu-Thr(PO4)-Gln-Glu.     

The scale-up from 0.1 to 100 liter of a unit process system based on 3-mm-diameter glass spheres for the production of four strains of FMDV from BHK monolayer cells

This paper describes the scale-up from 0.1 to 100 liter of the unit process based on 3-mm-diameter glass spheres for the growth of BHK monolayer cells. The production of four strains of FMD virus at the 0.1-, 10-, and 100-liter scales was examined. Cell growth was estimated from measurements of the concentration of glucose in the growth medium, while the release of virus was inferred from measuring the concentration of LDH in the culture supernatant fluid. The yields of virus at 0.1-, 1-, and 10-liter scales were similar but that from the 100-liter version was somewhat lower. The reason for this lower yield and the method used to overcome it over outlined.     

Relation of mechanical noise in the rat papillary muscle to the level of its contraction

The existence of mechanical noise (MN) has been demonstrated in isolated papillary muscles of rats at rest. The mean amplitude of the MN was about 1 mg, the mean frequency 1.5 Hz (t 22 degrees C). A good agreement was found between the MN amplitude and the contracture level of the muscle. However, during long contractures, the correlation between the noise and contracture magnitude was disturbed. There was no relationship between the MN amplitude and contracture magnitude during exposures inducing metabolic alterations (hypoxia, NaCN) and upsetting the work of the sarcoplasmic reticulum (caffeine). It is believed that the MN amplitude is in a good agreement with the contracture magnitude and, therefore, with the concentration of intracellular Ca2+, if the sarcoplasmic reticulum and contractile elements of the cells are intact.     

Possible incorporation of phosphoserine into globin readthrough protein via bovine opal suppressor phosphoseryl-tRNA

Suppressor [32P]phosphoseryl-tRNA, prepared using bovine seryl-tRNA synthetase and ATP:seryl-tRNA phosphotransferase, was mixed with rabbit reticulocyte lysates containing endogenous hemoglobin mRNA having the termination codon UGA (opal). The chromatographic pattern of the lysate on Sephacryl S-200 showed that the radioactivity of [32P]phosphate in the hot trichloroacetic acid-precipitate (phosphoprotein) was eluted at the position between mature hemoglobin and globin subunits. The phosphoprotein, obtained by chromatography on S-200, moved to the position corresponding to that of globin readthrough protein on SDS-PAGE. The analyses of the hydrolyzate of the phosphoprotein showed the presence of phosphoserine in the protein. These results suggest that animal opal suppressor tRNA functions in vitro to transfer phosphoserine to the position of the termination codon UGA (opal) on mRNA.     

Morphological taxonomy of little-differentiated Hyphomycetes

Morphological taxonomy of simple Hyphomycetes is complicated by the frequent occurrence of pleoanamorphism. In some groups of yeast-like fungi, uncommon synanamorphs are diagnostic. Differences in conidiogenesis do not always delimit natural groups. Some nomenclatural problems are mentioned, with an emphasis on the need of neotypification. Prospects are sketched for future taxonomic research.     

Binding of [3H]estradiol- and [3H]H1285-receptor complexes to rabbit uterine chromatin

In the present study we investigated the binding characteristics of estrogen and antiestrogen-receptor complexes to rabbit uterine chromatin. Activated or nonactivated estrogen receptors were partially purified by DEAE-cellulose chromatography using low (1 mM) or high (10 mM) concentrations of sodium molybdate. Activated [³H]estradiol-receptor complexes showed enhanced binding to chromatin acceptor sites unmasked by 1 M, 4 M and 6 M guanidine hydrochloride. We also examined the chromatin-binding characteristics of the estrogen receptors when bound by the high-affinity triphenylethylene antiestrogen, H1285. The acceptor site activity for the [³H]H1285-receptor complexes was markedly decreased at sites unmasked by 4 M and 6 M guanidine hydrochloride. Further, the nonactivated receptor complexes showed very low binding to deproteinized chromatin. The estrogen-receptor chromatin-acceptor sites were tissue specific and saturable. These chromatin acceptor sites differ in their affinity and capacity (number of binding sites per cell) for the estrogen- and antiestrogen-receptor complexes. Thus, we suggest that the differences in the physiological and physicochemical properties of estrogens and antiestrogens may be related to their differential interaction with uterine chromatin subfractions.     

Cloning and characterization of a SnRK1-encoding gene from <Emphasis Type="Italic">Malus hupehensis</Emphasis> Rehd. and heterologous expression in tomato

Sucrose non-fermenting-1-related protein kinase-1 (SnRK1) plays an important role in metabolic regulation in plant. To understand the molecular mechanism of amino acids and carbohydrate metabolism in Malus hupehensis Rehd. var. pinyiensis Jiang (Pingyi Tiancha, PYTC), a full-length cDNA clone encoding homologue of SnRK1 was isolated from PYTC by Rapid Amplification of cDNA Ends (RACE). The clone, designated as MhSnRK1, contains 2063 nucleotides with an open reading frame of 1548 nucleotides. The deduced 515 amino acids showed high identities with other plant SnRK1 genes. Quantitative real-time PCR analysis revealed this gene was expressed in roots, stems and leaves. Exposing seedlings to nitrate caused and initial decrease in expression of the MhSnRK1 gene in roots, leaves and stems in short term. Ectopic expression of MhSnRK1 in tomato mainly resulted in higher starch content in leaf and red-ripening fruit than wild-type plants. This result supports the hypothesis that overexpression of SnRK1 causes the accumulation of starch in plant cells. All the results suggest that MhSnRK1 may play important roles in carbohydrate and amino acid metabolisms.     

Primary production and accumulation of nutrients by the ground layer community of cerrado vegetation of central Brazil

Summary Net aerial primary production and accumulation of nutrients by the grasses and nongrasses of the ground layer community of a cerrado vegetation in central Brazil were determined in burnt and unburnt areas. The net aerial primary production of the ground layer community was 327 gm^–2 in the unburnt area and only 242 gm^–2 in the burnt area during the first year after fire. Grasses contributed 68 to 78% of the aerial biomass of the ground layer in the unburnt area. The live biomass in the burnt and unburnt areas was comparable by the end of the first dry period after the fire. The major part of N, P and K in the aerial biomass was in the grasses. The concentration of all nutrients in the aerial biomass was generally higher in the burnt area during the first year after the fire.