Chlorophyll and peptide compositions in the two photosystems of marine green algae

The molar ratios of chlorophyll a to b in the thalli of marine green algae were between 1.5 and 2.2, being appreciably lower than the ratio between 2.8 and 3.4 found for the leaves of higher plants and the cells of fresh-water green algae. The ratio of chlorophylls to P-700 in these marine algae was also lower than that in higher plants. The $\text{a}\text{b}$ ratios in the pigment proteins of Photosystems 1 and 2 separated by polyacrylamide-gel electrophoresis from sodium dodecyl sulfate-solubilized chloroplasts of four species of marine green algae, Bryopsis maxima, Cheatomorpha spiralis, Enteromorpha compress and Ulva conglobata, were approximately 5 and 1, which are considerably smaller than the ratios, 7 and 2, respectively, found for the pigment proteins of the two photosystems of higher plants separated by the same technique. The chloroplasts of Bryopsis maxima and Cheatomorpha spiralis lacked two of the peptides associated with Photosystem II, which are present in the chloroplasts of Spinacia oleracea and Taraxacum officinale.     

The ultrastructure of the ovarian follicle of medaka,Oryzias latipes

Summary The follicular cells in the oocytes of Oryzias latipes were studied by electron microscopy in order to clarify the fine structure, and the role of the cells during yolk formation and ovulation. The smallest follicles were observed during the early phase of peri-nucleolus stage of the oocyte. The cells have flattened nuclei, and perikarya with undeveloped organelles. But when the oocytes attain diameter of about 250 (yolk vesicle stage), both types of endoplasmic reticula are present. Moreover, the microvilli of the plasma membrane of oocyte as well as the follicles protrude into the pore canals of the zona radiata. In the oocytes of yolk stage the rough-surfaced endoplasmic-reticulum is typically developed and observed around the nuclei. Other organelles (lysosomes, mitochondria and Golgi) increase in number. The relation between the changes of cytoarchitecture in the follicles and yolk formation is discussed.At 17.00 p.m. on the day preceding ovulation the microvilli withdraw somewhat. Ribosomes are attached to the vesicular and cisternal endoplasmic reticula. When the oocytes attain complete maturation (24.00 p.m. at near ovulation), striking changes of the follicles are observed. The microvilli are almost withdrawn. In the degenerating follicles the lamellar structure is formed, and lipids are deposited at the center. At this time the contents of lysosomes have mostly disappeared.     

Effect of dopamine, bombesin and cysteamine hydrochloride on plasma growth hormone response to synthetic growth hormone-releasing factor in rats

In urethane anesthetized rats, an intracerebroventricular (icv) injection of 2 micrograms bombesin 5 min prior to the administration of synthetic human growth hormone-releasing factor (GRF) (1 microgram/kg, iv) inhibited plasma growth hormone (GH) response, while cysteamine hydrochloride (90 mg/kg, sc) administered 150 min beforehand depleted immunoreactive somatostatin content in the pituitary-stalk median eminence and consequently potentiated the response to GRF. Under the same experimental conditions, central injection of 1.89 micrograms (10(-8)M) dopamine hydrochloride or iv administration of L-DOPA (10 mg/kg) did not influence the subsequent plasma GH response to GRF. Results suggest indirectly that bombesin and cysteamine, but not dopamine, predominantly modulate somatostatin release from the hypothalamus.     

A mutagen precursor in Chinese cabbage, indole-3-acetonitrile, which becomes mutagenic on nitrite treatment

After treatment with nitrite, Chinese cabbage showed direct-acting mutagenicity on Salmonella typhimurium TA100 inducing 3100 revertants per g. One of the mutagen precursors that became mutagenic after nitrite treatment was isolated, and identified as indole-3-acetonitrile. After treatment with nitrite, 1 mg of indole-3-acetonitrile induced 17 400 revertants of TA100 and 21 000 revertants of TA98 without S9 mix.     

Analysis of receptor-binding site in Escherichia coli enterotoxin

Heat-labile enterotoxin produced by enterotoxigenic Escherichia coli and cholera enterotoxin are both composed of A and B subunits. The A subunit is an enzymatically active ADP-ribosylating subunit, while the B subunit, consisting of 103 amino acids, binds the toxin to a receptor, GM1-ganglioside, on the cell surface. A mutant isolated after treatment of E. coli producing heat-labile enterotoxin with N-methyl-N'-nitro-N-nitrosoguanidine produces a B subunit that is unable to bind to ganglioside. This subunit was purified and its primary amino acid sequence was determined. It differed from the native B subunit in only one amino acid at position 33; namely it had aspartate instead of glycine at position 33 from the N terminus. Thus glycine at position 33 from the N terminus of the B subunit is important for binding the B subunit to the ganglioside receptor.     

Charge heterogeneity of rat pituitary prolactin in relation to the estrous cycle, gonadectomy, and estrogen treatment

Pituitary samples were obtained from female rats at various stages of the estrous cycle, and from intact male and gonadectomized rats with and without estradiol treatment. The pituitary extracts with 60% EtOH pH 9.5, were fractionated by preparative isoelectric focusing (IEF), and immunoreactive prolactin (IR-PRL) was measured by RIA. Three types of IR-PRL molecular species were found in these IEF profiles. The first type (species A) was consistently found in an area of pH 4.5-5.4, and consisted of two main subspecies with pls 5.0 (Al) and 5.25 (A2). Species A occupied most part of pituitary IR-PRL in males, gonadectomized animals, and in females in a basal state such as diestrus (D) II 17:00. Species A was also found exclusively in the serum at proestrus (PE) 19:00. The amounts of species A decreased notably when the secretion became active from PE 15:00 to 22:00, then increased at estrus (E) 6:00 and 10:00 when the second type (species B), which was found in the area of pH 5.4-6.8 only in trace amounts at basal states, increased markedly. Species B decreased again at E 17:00, while species A fully recovered. Species B also increased when PRL biosynthesis was stimulated by estradiol in intact male and gonadectomized rats. These findings indicate that species A must be the storage and secretory type of IR-PR, and that species B must be IR-PRL in the biosynthetic process which is to be finally converted into species A. A third type (species C) was found in a region of pH 3.5-4.5 in the IEF profiles of gonadectomized animals. This species is possibly IR-PRL molecules under degradation. When the pituitary was extracted serially with 0.25 M ammonium sulfate pH 5.5 (fraction AMS) first, then with 60% EtOH pH 9.5 (fraction ET), fraction AMS contained mostly species B and C, while fraction ET contained species A almost exclusively. The results obtained with this differential extraction roughly coincided with IEF data, though some disagreements were observed.     

Conformational studies of myosin phosphorylated by protein kinase C

Smooth muscle myosin from chicken gizzard is phosphorylated by Ca2+-activated phospholipid-dependent protein kinase, protein kinase C, as well as by Ca2+/calmodulin-dependent kinase, myosin light chain kinase (Endo, T., Naka, M., and Hidaka, H. (1982) Biochem. Biophys. Res. Commun. 105, 942-948). We have now demonstrated the effect of phosphorylation by protein kinase C on the smooth muscle myosin molecule. In glycerol/urea polyacrylamide gel electrophoresis the 20,000-dalton light chain phosphorylated by protein kinase C co-migrated with that phosphorylated by myosin light chain kinase. Moreover, the light chain phosphorylated by both kinases migrated more rapidly than did the light chain phosphorylated by either myosin light chain kinase or protein kinase C alone. Myosin phosphorylated by protein kinase C formed a bent 10 S monomer while that phosphorylated by myosin light chain kinase was an unfolded and extended 6 S monomer in the presence of 0.2 M KCl. In addition, myosin phosphorylated by kinases had a sedimentation velocity of 7.3 S, thereby suggesting that the myosin was partially unfolded. The unfolded myosin was visualized electron microscopically. The fraction in the looped form was higher when for myosin phosphorylated by both kinases higher than for that phosphorylated by light chain kinase alone. Therefore, phosphorylation by protein kinase C does not lead to the change in myosin conformation seen with myosin light chain kinase.     

Temperature- and time-dependent inactivation of pyrroline-5-carboxylate synthase: suggestive evidence for an allosteric regulation of the enzyme

When pyrroline-5-carboxylate (PC) synthase activity in the membrane of mitochondria of rat small intestine mucosa was assayed in the presence of 0.5 mM ornithine, the time course of inactivation showed that the activity disappeared entirely by about 8 min at 30 degrees C, whereas there was no decrease in the activity at 15 degrees C. A prior incubation of the enzyme with ornithine at 30 or 37 degrees C in the presence of 50% sorbitol as a thermal stabilizer resulted in a marked loss of the activity, while that at 0 or 15 degrees C did not lose any. This suggests that PC synthase is inactivated by ornithine regardless of the presence of substrates. The inactivation at 30 degrees C proceeded gradually for about 7 h, until an equilibrium was attained. Extensive dialysis allowed the inactivated enzyme to regain about 60% of the original activity. These results suggest that the inactivation is reversible. The concentration of ornithine and the percentage of inactivation at equilibrium was correlated by the Hill equation and displayed a sigmoidicity with n = 1.47 and [S]50 = 0.036 mM. In the presence of sorbitol, the inactivation was prevented by 0.2 mM ATP or ADP. The role of the nucleotides in PC synthase regulation is discussed.