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161.
Klaus Rissler Peter Engelmann 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1996,679(1-2)
Conditions for the labeling of insulin with radioactive iodine isotopes were investigated by means of incorporation of non-radioactive 127I into the peptide. Either the chloramine-T (CT) or lactoperoxidase-hydrogen peroxide (LPO) technique was applied and reversed-phase high-performance liquid chromatography (RP-HPLC) was used for analysis of the reaction products. The LPO method provided the 127I-labeled peptide within 15–30 min, whereas the CT alternative yielded the labeled substrate even within 15 s. However, the latter reaction can only be controlled in a reproducible manner with difficulty and undesirad side-reactions became increasingly prominent when t a few seconds. In another experiment, the LPO technique was applied for radiolabeling insulin with 125I. The product was first purified by size-exclusion chromatography (SEC) and then subjected to RP-HPLC. SEC yielded two peaks. The smaller one, which eluted at a slightly higher Kd value (accounting for about 14% of total radioactivity) predominantly consisted of material eluting at the column's void volume under the conditions of RP-HPLC, whereas the main SEC fraction (accounting for about 86% of total radioactivity) yielded a single peak, as shown by HPLC. The radioactive material attributable to the main SEC fraction revealed the expected receptor-binding properties, as evidenced by displacement experiments with non-radioactive insulin, as well as the action of tetradecanoyl phorbol acetate on the binding characteristics and thus indicating formation of a labeled hormone retaining biological activity. 相似文献
162.
A receptor for protein import into potato mitochondria 总被引:3,自引:0,他引:3
Five potential surface receptors for protein import into plant mitochondria were identified by gentle trypsin treatment of intact mitochondria from potato tubers and subsequent preparation of outer mitochondrial membranes. One of them, a 23 kDa protein, was purified to homogeneity and analysed by direct protein sequencing. Copy DNA clones encoding the corresponding polypeptide were isolated with labelled oligonucleotides derived from the amino acid data. The 23 kDa protein shares significant sequence similarity with protein import receptors from fungal mitochondria and contains one of their typical tetratricopeptide motifs. Its integration into the outer membrane is independent of protease accessible surface receptors and not accompanied by proteolytic processing. Monospecific antibodies against the 23 kDa protein significantly reduce import capacity of isolated mitochondria indicating that this component is indeed involved in the recognition or import of precursor proteins. As in fungi, immunological inhibition of protein import with IgGs against a single receptor is incomplete suggesting the existence of other receptors in the outer mitochondrial membrane of plant mitochondria. 相似文献
163.
Klaus Werner Wolf Sabine Glatzel Frauke Niedereichholz Bryan M. Turner 《Biotechnic & histochemistry》1996,71(3):137-144
We describe a method for isolating chromosomes from testes of the desert locust, Schistocerca gregaria, and their subsequent incubation with antibodies directed against chromosomal proteins. The procedure involves hypotonic pretreatment of the germ cells, centrifugation onto coverslips in a cytocentrifuge and immunolabeling, while still unfixed, using a chromatin-stabilizing buffer. In the present case, an antibody specific for the acetylated isoforms of his tone H4 was tested. After the antibody treatment, the preparations are fixed using formaldehyde, stained with a DNA-specific fluorescent dye and mounted. Analysis of the preparations revealed good preservation of chromosome structure in prophase spermatogonia and late prophase I spermatocytes. Fully condensed chromosomes were not observed and are probably lost during preparation. The bright fluorescence of the autosomes indicates that the reaction between the antibody against acetylated histone H4 and its chromosomal antigen is not impeded. In contrast, the X univalent remained unstained with the exception of a small terminal band. Thus, cytospin preparations of locust germ cells allow high resolution immunolabeling with antibodies against chromosome-associated proteins. 相似文献
164.
Klaus Dose 《Journal of biological physics》1995,20(1-4):181-192
165.
The present study was conducted to investigate the effect of zinc deficiency on fatty acid desaturation in rats fed two different
types of dietary fat, a mixture of coconut oil and safflower oil (7∶1, w/w, “coconut oil diet”) or linseed oil (“linseed oil
diet”). In order to ensure an adequate food intake, all rats were force-fed by gastric tube. Zinc deficiency caused statistical
significant reducion of Δ9-desaturase activity in liver microsomes of rats fed coconut oil diet and tendencial reduction (p<0.15) in rats fed linseed oil diet compared with control rats fed diets with the same type of fat. In agreement with this
effect, zinc deficiency in the rats fed both types of dietary fat increased the ratio between total saturated and total monounsaturated
fatty in liver phospholipids and liver microsomes. Zinc deficient rats on the coconut oil diet had unchanged Δ6-desaturase
activity with linoleic acid as substrate and lowered activity with α-linolenic acid as substrate. In contrast, zinc deficient
rats on the linseed oil diet had increased Δ6-desaturase activity with linoleic acid as substrate and unchanged activity with
α-linolenic acid. Because linoleic acid is the main substrate for Δ6-desaturase in the rats fed coconut oil diet, and α-linolenic
acid is the main substrate in the rats fed linseed oil diet, it is concluded that in vivo Δ6-desaturation was not changed
by zinc deficiency in the rats fed both types of dietary fat. Activity of Δ5-desaturase was also not changed by zinc deficiency
in the rats fed both dietary fats. Levels of fatty acids in liver phospholipids and microsomes derived by Δ4-, Δ5-, and Δ6-desaturation
were not consistently changed by zinc deficiency in the rats fed both types of dietary fat. Thus, the enzyme studies and also
fatty acid composition data of liver phospholipids and microsomes indicate that zinc deficiency does not considerably disturb
desaturation of linoleic and α-linolenic acid. Therefore, it is suggested that similarities between deficiencies of zinc and
essential fatty acids described in literature are not due to disturbed desaturation of linoleic acid in zinc deficiency. The
present study also indicates that zinc deficiency enhances incorporation of eicosapentaenoic acid into phosphatidylcholine
of rats fed diets with large amounts ofn-3 polyunsaturated fatty acids. 相似文献
166.
167.
Klaus Hövemeyer 《Oecologia》1995,102(1):86-94
The food web centering on Allium ursinum (Liliaceae) in a beech forest (Germany) is described, and temporal variation of active trophic links is related to species' life cycles. The most important insect herbivores are Cheilosia fasciata (a larval leaf miner) and Portevinia maculata (a larva bulb miner) (Diptera: Syrphidae). Energy, carbon and nitrogen flow in the food chain (Allium-Cheilosia-Phygadeuon ursini) are investigated and analysed with respect to differences in resource allocation by the leaf miner and its hymenoptereous parasitoid. In C. fasciata nitrogen is likely to be the limiting resource, while growth in Phygadeuon ursini appears energy-limited. Larval feeding habits of C. fasciata and Portevinia maculata determined the timing of the species' life cycles and, as a consequence, appeared to preclude the existence of a pupal parasitoid in Portevinia maculata. Further details of life history traits are demonstrated and discussed. 相似文献
168.
Lars Berglund Janne Brunstedt Klaus K. Nielsen Zhaochun Chen Jørn D. Mikkelsen Kjeld A. Marcker 《Plant molecular biology》1995,27(1):211-216
A gene (Chl) encoding a novel type of chitinase was isolated from Beta vulgaris. The Ch1 protein consists of an N-terminal hydrophobic prepeptide of 25 amino acids followed by a hevein-like domain of 22 amino acid residues, an unusually long proline-rich domain of 131 amino acid residues with 90 prolines, and finally a catalytic domain of 261 amino acid residues. Proteins with similar proline-rich domains are present in some other plants. The Chl gene shows a transient expression in response to fungal infection. 相似文献
169.
the entire mitochondrial genome (mt genome) of the unicellular green alga Platymonas subcordiformis (synonym Tetraselmis subcordiformis; Prasinophyceae) was cloned and a physical map for the four restriction enzymes Hind III, Eco RI, Bgl II and Xba I was constructed. The mt genome of P. subcordiformis is a 42.8 kb circular molecule, coding for at least 23 genes. Hybridization and sequence analysis revealed the presence of a ca. 1.5 kb inverted repeat on the mt genome of P. subcordiformis. Phylogenetic analyses based on sequences of several coxI genes were carried out. Our data indicate that mitochondria from P. subcordiformis and from land plants form a natural, monophyletic group. 相似文献
170.
Prof. Dr. Wolf-Christian Dullo Dr. Marcos Gektidis Prof. Dr. Stjepko Golubic Dr. Georg A. Heiss Dipl. Biol. Heike Kampmann Dr. William Kiene Dipl. Ökol. Dieter K. Kroll Dipl. Biol. Martin L. Kuhrau Dr. Gudrun Radtke Dr. John G. Reijmer Dr. Götz B. Reinicke Prof. Dr. Dietrich Schlichter Prof. Dr. Helmut Schuhmacher Klaus Vogel 《Facies》1995,32(1):145-188