Genetic identification of rat liver carboxylesterases isolated in different laboratories

Six carboxylesterases previously isolated from rat liver microsomes, characterized in Brussels and in Kiel, were compared with genetically defined liver esterases of various reference strains using polyacrylamide gel electrophoresis and isoelectric focusing. The six liver carboxylesterases were identified as alloenzymic forms of ES-3, ES-4, ES-8/ES-10 and ES-15 according to the genetic nomenclature recommended by van Zutphen (Van Zutphen, L.F.M. (1983) Transplant. Proceed. 15, 1687-1688). The genetic and biochemical characteristics of the four isoenzymes are summarized, and their identity with several other drug-metabolizing esterases/amidases and lipases of rat liver endoplasmic reticulum is discussed.     

tRNA recognition site of Escherichia coli methionyl-tRNA synthetase

We have previously shown that anticodon bases are essential for specific recognition of tRNA substrates by Escherichia coli methionyl-tRNA synthetase (MetRS) [Schulman, L. H., & Pelka, H. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 6755-6759] and that the enzyme tightly binds to C34 at the wobble position of E. coli initiator methionine tRNA (tRNAfMet) [Pelka, H., & Schulman, L. H. (1986) Biochemistry 25, 4450-4456]. We have also previously demonstrated that an affinity labeling derivative of tRNAfMet can be quantitatively cross-linked to the tRNA binding site of MetRS [Valenzuela, D., & Schulman, L. H. (1986) Biochemistry 25, 4555-4561]. Here, we have determined the site in MetRS which is cross-linked to the anticodon of tRNAfMet, as well as the location of four additional cross-links. Only a single peptide, containing Lys465, is covalently coupled to C34, indicating that the recognition site for the anticodon is close to this sequence in the three-dimensional structure of MetRS. The D loop at one corner of the tRNA molecule is cross-linked to three peptides, containing Lys402, Lys439, and Lys596. The 5' terminus of the tRNA is cross-linked to Lys640, near the carboxy terminus of the enzyme. Since the 3' end of tRNAfMet is positioned close to the active site in the N-terminal domain [Hountondji, C., Blanquet, S., & Lederer, F. (1985) Biochemistry 24, 1175-1180], this result indicates that the carboxy ends of the two polypeptide chains of native dimeric MetRS are folded back toward the N-terminal domain of each subunit.     

GC-MS identification of GA20-13-O-glucoside formed from GA20 in normal plants and dwarf-1 mutants ofZea mays L.

After feeding GA₂₀ to excised seedlings ofZea mays L. normals (N) and dwarf-1 mutants (d1), GA₂₀-13-O-glucoside (9) was identified by HPLC and by GC-MS of its permethylated derivative. The glucosylation rate of GA₂₀ was found to be higher in the dwarf-1 mutant (26%) than in the normal plant (3.6%). This article includes a GC-MS study in which diagnostic fragments from the spectra of permethylated synthetic GA glucosides have been selected that proved to be useful for the identification of permethylated GA glucosides.     

Comparative gastrointestinal morphology of the Kori bustard and secretary bird

Two secretary birds and three Kori bustards were studied to determine differences between their body size and gastrointestinal morphology. Body measurements were made on captive, live birds and gastrointestinal measurements on fresh postmortem specimens. For predator species, such as the Kori bustard and secretary bird, body size is a function of their ability to capture and destroy prey. While the secretary bird was clearly the taller of the two species, superior body weight, wing length, and therefore body size was noted for the Kori bustard. The size and length of the gastrointestinal tract varied between species. The secretary bird had the shorter, less complex digestive tract, with a foregut well adapted for consumption of large quantities of flesh. The large intestine was devoid of ceca. The gastrointestinal tract of the Kori bustard was markedly different from that of the secretary bird. The foregut was less complex and the large intestine possessed large, voluminous ceca.     

Immunolocalization of alpha and beta chains of human chorionic gonadotropin,placental lactogen and pregnancy-specific beta-glycoprotein in term placenta by a touch preparation method

Summary Touch preparations of human placenta yield cells retaining antigenic reactivity to immunoperoxidase stains for and chains of human chorionic gonadotropin, placental lactogen, and pregnancy-specific glycoprotein. This method is a rapid and simple alternative to conventional frozen and paraffin-embedded sections for detection of placental peptides.     

Plasmid-encoded copper resistance and precipitation by Mycobacterium scrofulaceum.

A copper-tolerant Mycobacterium scrofulaceum strain was able to remove copper from culture medium by sulfate-dependent precipitation as copper sulfide. Such precipitation of copper sulfide was not observed in a derivative that lacks a 173-kilobase plasmid. In addition, the plasmid-carrying strain has a sulfate-independent copper resistance mechanism.     

Effect of nutritional insufficiency on cell proliferation in the matrix zones of the cerebellar anlage in mice

The work has been performed on 62 CBA mice. In the ventricular zone and in the external granular layer of the cerebellar anlage of embryos (13-17 days of the intrauterine development) mitotic index, labelled nuclei index, part of labelled mitoses have been counted. Parameters of the mitotic cycle of the matrix cells have been calculated by means of the graphic method. The proliferative pool value has been calculated. At malnutrition the cerebellar anlage structure retards in its maturation from the norm. For the matrix zones of the cerebellar anlage, higher indices of the proliferative activity are specific. At the same time, duration of the mitotic cycle of the matrix cells increases by 15-17%. It is possible, that retardation of histogenesis of the mouse cerebellar anlage, when developing under conditions of alimentary insufficiency depends on decreased rate of cell proliferation, as a result of prolonged mitotic cycle of the matrix cells.     

Effect of beta-hydroxybutyrate and acetoacetate on insulin and glucagon secretion from perfused rat pancreas

To elucidate the physiological significance of ketone bodies on insulin and glucagon secretion, the direct effects of beta-hydroxybutyrate (BOHB) and acetoacetate (AcAc) infusion on insulin and glucagon release from perfused rat pancreas were investigated. The BOHB or AcAc was administered at concentrations of 10, 1, or 0.1 mM for 30 min at 4.0 ml/min. High-concentration infusions of BOHB and AcAc (10 mM) produced significant increases in insulin release in the presence of 4.4 mM glucose, but low-concentration infusions of BOHB and AcAc (1 and 0.1 mM) caused no significant changes in insulin secretion from perfused rat pancreas. BOHB (10, 1, and 0.1 mM) and AcAc (10 and 1 mM) infusion significantly inhibited glucagon secretion from perfused rat pancreas. These results suggest that physiological concentrations of ketone bodies have no direct effect on insulin release but have a direct inhibitory effect on glucagon secretion from perfused rat pancreas.     

The isolation and characterisation of a cDNA clone for human lecithin:cholesterol acyl transferase and its use to analyse the genes in patients with LCAT deficiency and fish eye disease

We have isolated cDNA clones coding for human lecithin:cholesterol acyl transferase (LCAT) from a liver-specific cDNA library by the use of two oligonucleotide probes based on the protein sequence. The clones span the sequence coding for the entire secreted LCAT, the 3' untranslated sequence and 12 amino acids of the signal peptide. The peptide sequence contains the conserved active site of serine lipases within a hydrophobic domain, flanked by a possible amphipatic alpha-helix. Only one gene for LCAT could be detected in genomic blots. We have used the cDNA as a probe to analyse the LCAT gene in patients suffering from LCAT deficiency and fish eye disease. No rearrangements or abnormal gene fragments were detected in these patients.     

High affinity binding of human interleukin 4 to cell lines

Purified human recombinant interleukin 4 (IL-4) was radio iodinated to high specific radioactivity without loss of biological activity. 125I-IL-4 bound specifically to the Burkitt lymphoma Jijoye cells and other cell lines. Jijoye cells showed a high affinity for 125I-IL-4 (Kd approximately equal to 7 10(-11) M) and displayed 1200-1400 specific receptors per cell at 4 degrees C or 37 degrees C. The equilibrium dissociation constant (Kd) corresponds to the IL-4 concentration which induces 50% maximal expression of the low affinity IgE receptor (Fc epsilon RL/CD23) on Jijoye cells. At 4 degrees C the rate constant of association K1 is 1.7 x 10(6) M-1 s-1 and the rate contant of dissociation k -1 is 1.3 x 10(-4) s-1 (t 1/2 = 91 min.) No human recombinant lymphokines other than IL-4 were able to compete for the binding of 125I-IL-4 to its receptor.