Studies on the subunit structure and amino acid sequence of triose phosphate isomerase from chicken breast muscle

1. Triose phosphate isomerase was prepared by chromatography on DEAE-cellulose of an (NH(4))(2)SO(4) fraction of an extract of homogenized chicken breast muscle. The product is homogeneous on gel electrophoresis and is suitable for growing crystals for X-ray work. The specific activity is 10000 units/mg and the value for E(0.1%) (280) is 1.20. 2. Comparison between the sum of the amino acid compositions of the tryptic peptides of the protein and the amino acid composition obtained on total hydrolysis of the protein indicates that the relative subunit mass is about 27000. 3. These data, together with the results of the examination of the amino acid compositions of a number of minor peptides, the number of peptides in the tryptic digest and the complete amino acid sequences of the tryptic peptides (the determination of which is described here), give no indication that the subunits are dissimilar. 4. A tentative amino acid sequence is presented for the protein, in which the ordering of the tryptic peptides is derived by homology with the sequence of the rabbit muscle enzyme (Corran & Waley, 1973). 5. An appendix describes the use that was made of mass spectrometry in the determination of some of the sequences. Mass-spectrometric data have been obtained for 35 residues, that is about 15% of the total sequence of the protein. 6. An extended version of the present paper has been deposited as Supplementary Publication SUP 50025 at the British Library, Lending Division (formerly the National Lending Library for Science and Technology), Boston Spa, Yorks. LS23 7BQ, U.K., from whom copies may be obtained on the terms given in Biochem. J. (1973) 131, 5.     

The problem of carotenoid biosynthesis in the taxonomy of genera Rhodotorula and Rhodosporidium

This report presents results in the composition of major carotenoids of various coloured mutants of the genusRhodotorula and of mating types of the genusRhodosporidium. The separation of carotenoid intermediates was carried out by thin layer chromatography using Silufol 254 Kavalier and the determination of eluated spots by spectrophotometry. There was found no difference in carotenoid composition of both mating typesa and α of individual species of the genusRhodosporidium. The vegetative and sexual reproduction ofRhodotorula andRhodosporidium can be separated from the carotenogenesis using 10^?4 mol diphenylamine. It was concluded that lycopene could be the intermediate to mono- and dicyclic carotenoids; in the case of partial inhibition of the dehydrogenation step the direct cyclization of neurosporene to β-zeacarotene can be expected. An unknown compound, probably lycopersene was found and was considered to be the precursor of phytoene. Phytoene and phytofluene were proved in all studied samples. Nutritional conditions (vitamins, sulfur amino acids, etc.) are able to shift the ratios between major carotenoids. Rhodotorula aurantiaca strains were observed to be auxotrophic mutants of various characters and the existence of this species as independent one, was denied.     

Macrophage regulation of DNA synthesis in lymphoid cells: effects of a soluble factor from macrophages

Addition of rat peritoneal macrophages to nonadherent rat spleen cells in culture results in enhancement or suppression of DNA synthesis depending on the ratio of macrophages to lymphocytes. At high ratios of macrophages to lymphocytes (1:5), suppression can be observed as early as four hours. Macrophages suppress incorporation of thymidine (TdR) by nonadherent spleen, thymus and bone marrow cells, in most instances, to less than 5% of that observed in culture to which macrophages were not added. In the presence of macrophages, incorporation of [³H]uridine and [¹⁴C] amino acids by spleen cells was also moderately suppressed. Based on ⁵¹Chromium release and dye exclusion assays, it appears that suppression is not due to cytotoxicity. Furthermore, suppression of [³H]TdR incorporation by nonadherent spleen cells is reversible, in the presence of an antigenic stimulus, following removal of the macrophages from the cultures. The suppressive effects are not elicited by extracts of macrophages, freeze-thawed or heated macrophages, but appear to be due to a low molecular weight, heat stable factor released into the macrophage culture fluid.     

Sterols in species of Pythium

Summary Analysis by gas chromatography revealed the presence of small amounts of squalene, but not lanosterol nor ergosterol in Pythium paroecandrum, P. ultimum, P. graminicola, and P. arrhenomonas. However, when acetate-¹⁴C was used as a precursor for sterols, even squalene was not found in P. graminicola. The deficiency in the sterol synthesizing mechanism may therefore be at or before the squalene forming step. Both squalene and ergosterol were present in the mycelium of Rhizoctonia solani, as shown by both gas chromatography and by the incorporation of acetate-¹⁴C into ergosterol. The absence of ergosterol in Pythium and its presence in Rhizoctonia is consistant with the resistance to the antibiotic filipin of Pythium species and the sensitivity of R. solani.     

Bemerkungen zur Taxonomie vonVicia sepium L.

In der ArtVicia sepium L. kann man in Mitteleuropa und in südlichen Teilen von Nordeuropa drei Varietäten unterscheiden: 1. var.sepium mit breiten Blättchen und einem kahlen bis angedrückt behaarten Kelch, 2. var.montana Koch mit schmalen Blättchen und einem kahlen oder zerstreut angedrückt behaarten Kelch, und 3. var.eriocalyx ?elak. mit breiten bis mittelbreiten Blättchen und einem abstehend langbehaarten Kelch, noch mit einer niedrigen und einer hohen Wuchsform. Bei allen Proben wurde die Chromosomezahl 2n=14 festgestellt.     

Bemerkungen zur Taxonomie und Nomenklatur vonLotus krylovii Schischk. etSerg. undL. corniculatus L. subsp.frondosus Freyn

Lotus krylovii Schischk. etSerg. undL. corniculatus L. subsp.frondosus Freyn sind zwei unterschiedliche Taxa, gekennzeichnet durch einige morphologische Merkmale.L. krylovii ist am nächsten mitL. tenuis Waldst. etKit. verwandt;L. corniculatus subsp.frondosus gehört in den Umkreis der Unterart vonL. corniculatus L., die im östlichen Teil des Areals der Art verbreitet ist. Nach ihrer Haupt-Verbreitung gehören die beiden Taxa zu den westasiatischen Arten.     

The effect of sterols on the metabolism of Pythium species

Summary The growth of several Pythium species is increased between 65 and 100% if cholesterol is added to the growth medium. The optimum concentration is 15 mcg per ml. Mycelium of Pythium ultimum, in which cholesterol is present, incorporates glucose-U-¹⁴C and releases ¹⁴CO₂ at a faster rate than the corresponding sterol free mycelium. In sterol containing cells, more ¹⁴CO₂ is produced from a given amount of absorbed glucose-U-¹⁴C than in sterol free cells, there is thus in sterol containing hyphae a higher level of energy production. This condition can account for the increase in growth due to cholesterol. Only if sterols are present in the cellular membranes of Pythium species is the optimum synthetic capacity reached.     

The genusSaccharomyces (Meyen) Reess

The authors submit a taxonomic evaluation of an intermediate group of strains between the speciesSaccharomyces carlsbergensis Hansen andSaccharomyces cerevisiae Hansen. The material consisted of atypical strains of “bottom” brewer’s yeasts and the synonymous strainsSaccharomyces monacensis Hansen andSaccharomyces mandshuricus Saito. It was found that there were two different serological types in the speciesSaccharomyces carlsbergensis, one of which was characterized by the presence of antigen “C” and was typical for this species, while the other possessed antigen “M” and was grouped roundSaccharomyces monacensis. This second serological type merges with a group of strains which gives only one third fermentation of raffinose, so that it is actually an intermediate betweenSaccharomyces cerevisiae Hansen andSaccharomyces carlsbergensis Hansen and indicates the course of progressive development from the former species to the latter. No close similarity was found betweenSaccharomyces mandshuricus Saito and some of the strains of the transitional group or typical representatives of the two main species, and the authors therefore consider that there is some obscurity as to its synonymity withSaccharomyces carlsbergensis.