Polyamine Distribution Profiles within the Phyla Nematoda, Platyhelminthes, Annelida and Cnidaria

Cellular polyamines of nematodes of the phylum Nematoda, planarians, cestodes and trematodes of the phylum Platyhelminthes, earthworms, leeches and sandworms of the phylum Annelida, and hydras of the phylum Cnidaria, were analyzed by HPLC and GC to obtain phylogenetic informations and parasitic peculiarities on polyamine profiles in lower invertevrates. Spermidine and spermine were ubiquitously distributed in four zoo-parasitic nematodes. A plant parasite, Pratylenchus and a fungivore, Bursaphelenchus and two entomopathogens, Steinernema as well as free-living bacteriovores, Caenorhabditis and Dorylaimus, contained spermidine and lacked spermine. Zoo-parasitic cestodes Hymenolepis and Diphyllobothrium, and zoo-parasitic trematodes Paragonimus, Fasciola and Schistosoma, belonging to the phylum Platyhelminthes, ubiquitously contained spermidine and spermine, suggesting their dependence on host animals for uptake of spermine. Putrescine, cadaverine, spermidine and spermine were the major polyamines in freshwater, marine and land planarians (the phylum Platyhelminthes), and free-living earthworms, leeches and sandworms (the phylum Annelida). Hydras belonging to the phylum Cnidaria contained putrescine and spermidine alone. Homospermidine was distributed in a zoo-parasitic helminth of the phylum Nematoda and two zoo-parasitic tapeworms of the phylum Platyhelminthes as well as planarians, earthworms and leeches. Norspermidine and/or norspermine were found in two land planarians, a leech and a sandworm. This is the first report on the occurrence of these novel polyamines within lower invertebrates

Effect of the Degree of Cross-Linkage of Polyacrylamide Gel on the Electrophoretic Mobility of Histones

The electrophoretic mobility in polyacrylamide gel of calf thymus histones was examined as a function of the degree of cross-linkage (C,%) of the gel in five different buffer systems. With increasing the degree of cross-linkage, the mobility once decreased and then increased, irrespective of total gel concentrations (T,%) used (5-15%). The degree of cross-linkage of the gel where the mobilities of the histones became minimum (C_), 7-15%C, was larger than that where the mobilities of other high molecular weight proteins become minimum, 5-7%C. The retardation coefficients (K_R) of the histones obtained from changes of the total gel concentration attained their maximum values in gels which have C_. In gels of C_ an approximately linear relationship was obtained between the K_R and the molecular weight of the histones

DNA Strand Scission by Mitoxantrone and Binding of Anti-Cancer Quinones to Chromatin DNA

The λ phage DNA and hog thyroid DNA after exposure to a hydroxyl radical-generating system containing NADPH-cytochrome P-450 reductase and anthraquinones such as mitoxantrone (CL 232315) and CL 232468 lacked in intact DNA fragments, indicating DNA strand breaks. The mechanism of DNA scission by the drugs closely resembles that induced by adriamycin and mitomycin C. When isolated hog thyroid chromatin was exposed to the system containing mitoxantrone, fragmentation of chromatin DNA was not detected. DNase I digestion for chromatin DNA was inhibited by the presence of the anthraquinones, suggesting intercalation of the drugs to DNase I-sensitive DNA regions in chromatin without DNA strand breaks

Effect of Temperature on Polyacrylamide Gel Electrophoresis and Chromatography of Histones and Protamines

Polyacrylamide gel electrophoresis or chromatography of histories (H1, H5, H2A, H2B, H3 and H4) and protamines (salmine and clupeine) in 0.9M acetic acid or 0.01M HC1, respectively, was carried out at various temperatures. A retardation of either electrophoretic migration or chromatographic elution of the proteins was observed at lower temperatures. H1 and H5 histones exhibited the least extent of retardation. The retardation with lowering temperature was also observed in the presence of 0.02-0.05M NaCl but not in the presence of 5-6.25M urea. The extent of retardation in polyacrylamide gel chromatography with lowering temperature increased with proteins of higher arginine contents. The fact was explained as that ionic interactions between guanido groups of arginine residues in the proteins and carboxyl groups of polyacrylamide are strengthened at lower gel temperatures

Polyamine Distribution Patterns in Coryneform Bacteria and Related Gram-Positive Eubacteria

Polyamines of the nine genera of coryneform and related Gram-positive eubacteria were analyzed by HPLC. Authentic species of Microbacterium, Aureobacterium, Cellulomonas and Corynebacterium were devoid of polyamines. Arthrobacter species were divided into polyamine-absent, putrescine, cadaverine and putrescine-cadaverine types. Clavibacter contained putrescine and cadaverine. Spermidine was detected in some species of Brevibacterium, Exiguobacterium and Curtobacterium and diaminopropane in some Curtobacterium. Heterogeneous polyamine profiles were found in the three genera

Triton-Gel Electrophoresis of Histones Containing Oxidized Methionine and Cysteine Residues

Electrophoresis in polyacrylamide gels in the presence of 6mM Triton X-100 in 0.9M acetic acid and 6.25M urea gave a high resolution of separation of the histones in which methionines and cysteines, respectively, are oxidized to the sulfoxides or sulfones (H2A, H2B, H3, H4) and to form an intramolecular disulfide bond (H3). The methionine-oxidized histones migrated faster than untreated histones. The electrophoretic mobility of histone H3 also increased by oxidation of the two cysteines to a cystine. The increase of the mobilities might be explained in terms of the decrease of the helical contents of the histones by the oxidation of methionine and/or cysteine residues