Axial ligands of chloroplast cytochrome b-559: identification and requirement for a heme-cross-linked polypeptide structure

Optical, resonance Raman, and electron paramagnetic resonance spectroscopies have been used to characterize the ligands and spin state of the chloroplast cytochrome b-559. The protein was isolated from both maize and spinach in a low-potential form. The spectroscopic data indicate that the heme iron in both ferric and ferrous cytochrome b-559 is in its low-spin state and ligated in its fifth and sixth coordination positions by histidine nitrogens. Electron paramagnetic resonance data for the purified spinach cytochrome are in good agreement with those determined by Bergstr?m and V?nng?rd [Bergstr?m, J., & V?nng?rd, T. (1982) Biochim. Biophys. Acta 682, 452-456] for a low-potential membrane-bound form of cytochrome b-559. The g values of high-potential cytochrome b-559 are shifted from those of its low-potential forms; this shift is interpreted as arising from a deviation of the planes of the two axial histidine imidazole rings from a parallel orientation. The model is consistent with the physical data and may also account for the facility with which cytochrome b-559 can be converted between low- and high-potential forms. Recent biochemical and molecular biological data [Widger, W. R., Cramer, W. A., Hermodson, M., Meyer, D., & Gullifor, M. (1984) J. Biol. Chem. 259, 3870-3876; Herrmann, R. G., Alt, J., Schiller, D., Cramer, W. A., & Widger, W. R. (1984) FEBS Lett. 179, 239-244] have shown that two polypeptides, one with 83 residues and a second with 39 residues, most likely constitute the protein of the cytochrome.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparative study of histones from slime mold Physarum polycephalum and calf thymus

The histones from slime mold Physarum polycephalum and calf thymus were characterized in terms of some physico-chemical properties. The molecular weights of six principal histone fractions of Ph. polycephalum were found to be the following: P1--22 700, P3--15 700, P4a--15 000, P4b--14 300, P5--12 800 and P6--10 500. Electrophoretically homogenous histone fractions H1, H2b and H4 of calf thymus and histones P1, P3, P4b and P6 of slime mold were obtained by gel-filtration on Acrylex P-60. These findings suggest that fractions P1, P4a, P4b, P5 and P6 of slime mold Ph. polycephalum are homologus with respect to the histone fractions H1, H3, H2b, H2a and H4 of calf thymus. Only fraction P3 has no corresponding fraction in the calf thymus histones; a fraction corresponding to histone P3 of slime mold was absent.     

NUTRITION OF TRIDACHIA CRISPATA (MORCH) (SACOGLOSSA)

Autotrophic nutrition plays an important role in adult Tridachiacrispata in the western Atlantic Ocean and results from an endosymbioticassociation between the sacoglossan and the kleptoplastids ofChlorophyta. A series of living specimens 7-80 mm in length was studied inthe laboratory. After a body length of 10 mm has been attained,the parapodial lobes of the two sides meet and fuse. Microscopicexamination of faecal strings of specimens undergoing this transitionrevealed that algal remains were produced only by juvenilesless than 13 mm long. Larger specimens defaecated only rarelyand the diminutive faecal strings contained no recognizablealgal debris. This transition probably parallels the physiologicaltransition between juvenile heterotrophic nutrition and adultautotrophic nutrition. Microscopic examination of the radulae of transitional specimensshowed no diminution of the size, number or morphology of theteeth. This indicates that in an emergency heterotrophic nutritionmay be resumed. Growth rates of Tridachia were measured in the laboratory, withand without light and the provision of algal foods. Unfed animalslost weight rather rapidly in captivity, whereas those fed uponsuitable algae (Halimeda discoidea, Caulerpa verticillata, C.racemosa and Chaetomorpha sp) did not.Caulerpa sertularioideswas suitable for food for a limited time, but proved toxic afterabout a week. Of the other species tested, Caulerpa verticillatawas the least suitable, while Halimeda, Chaetomorpha and Caulerparacemosa were the best. Ironically, C. verticillata was theonly one of the algal species tested which is known to be takenin the natural diet in Floridean waters. These experiments confirmedthat heterotrophic feeding will be resumed in Tridachia if theculture-conditions do not favour autotrophic nutrition.     

Commitment to Ploidy Conversion of 3y1 Cells During Metaphase Arrest By Colcemid

Abstract. Diploid rat 3Y1 fibroblasts proliferate to a saturation density, where they are arrested with a 2N DNA content. After treatment to induce ploidy conversion, the conversion rate can be estimated by determining the fraction of cells with a 4N DNA content in the confluent culture using flow cytometry. Using this method it was found that during mitotic inhibition with colcemid, 3Y1 cells were converted to tetraploids with a high efficiency (above 80%); the optimum colcemid concentration and exposure period were 40 ng/ml and 8 hr, respectively. When metaphase cells were reseeded with 40 ng/ml of colcemid, they delayed anchorage to a dish; 6 hr was required for complete adhesion (in the absence of colcemid only 1 hr was required). When reseeded metaphase cells were exposed to 40 ng/ml of colcemid for 5 hr followed by its removal, a greater fraction of the cells anchored to the substratum were converted to tetraploids, whereas most of the floating cells were not. A greater fraction of the anchored cells had formed nuclei, whereas most of the floating cells preserved condensed metaphase chromosomes. These results indicate that the cells which have formed nuclear structure without chromosome separation during mitotic inhibition are irreversibly committed to ploidy conversion, with restoration of anchorage.     

The stimulation of the phosphorylation of ribosomal protein S6 by cycloheximide and puromycin

The administration of cycloheximide or puromycin to rats in amounts that all but completely inhibited hepatic protein synthesis caused an increase in the amount of radioactive phosphate incorporated into the liver ribosomal protein S6; there was also an increase in the prominence of the derivatives of S6 which contain increasing numbers of phosphorylated serine residues.