Structural and functional relationships of human ferritin H and L chains deduced from cDNA clones

We have isolated essentially full-length cDNA clones for human ferritin H and L chains from a human liver cDNA library. This allows the first comparison of H and L nucleotide and amino acid sequences from the same species as well as ferritin L cDNA sequences from different species. We conclude that human H and L ferritins are related proteins which diverged about the time of evolution of birds and mammals. We also deduce the secondary structure of the H and L subunits and compare this with the known structure of horse spleen ferritin. We find that residues involved in subunit interaction in shell assembly are highly conserved in H and L sequences. However, we find several interesting differences in H subunits at the amino acid residues involved in iron transport and deposition. These substitutions could account for known differences in the uptake, storage, and release of iron from isoferritins of different subunit composition.     

A plasmodial alpha-tubulin cDNA from Physarum polycephalum. Nucleotide sequence and comparative analysis

As the first step towards correlating structure and function of tubulin in the slime mold Physarum polycephalum we have elucidated the nucleotide sequence of a cDNA that appears to code for all but the last 25 to 30 C-terminal amino acids of a plasmodial alpha-tubulin. Differences in amino acid sequence from those of other alpha-tubulins are distributed fairly evenly throughout the sequence, although a relatively extensive conserved region is found in position 396 to 426 near the C terminus. A small region in position 298 to 307 contains a cluster of amino acid residues unique to Physarum alpha-tubulin. The sequence is 70% homologous to two yeast alpha-tubulins and about 83% homologous to five animal alpha-tubulins. A comparison of the homologies of all the known alpha-tubulins indicates that a large decrease in the accepted point mutation rate has occurred during the evolution of the metazoa, suggesting a major functional specialization of microtubules.     

Effect of acid or enzymatic hydrolysis on ethanol production by Zymomonas mobilis growing on Jerusalem artichoke juice

Summary Ethanol production from the inulin of Jerusalem artichoke byZ. mobilis was studied in batch and continuous fermentations. Both acid or enzymatic hydrolysis were used. In continuous cultures enzymatic hydrolysis showed better results. Ethanol productivities of 17.7 and 29.0 g/l.h were obtained at output concentrationsca 35 g/l (% of conversion 99 and 83; ethanol yield 0.45 g/g). The hydrolysed juice could be used without any nutrient addition.     

Identification and isolation of the primary aggregation factor from the cell membrane of the sponge Geodia cydonium

Summary The primary aggregation factor (pAF) of sponge cells is a glycoprotein that is firmly associated with the cell membrane. Polyspecific antibodies (anti-GM) prepared from sera raised against membranes of cells from the siliceous sponge Geodia cydonium were found to inhibit initial aggregation of homologous cells. The inhibition of aggregation, caused by anti-GM was neutralized by pAF. The pAF had been successfully solubilized and enriched by affinity chromatography, gel filtration and density gradient centrifugation, if checked by polyacrylamide gel electrophoresis in the presence of urea. The M_r of the native pAF was approximately 40 000 as estimated by gel filtration; under denaturing conditions three protein species (M_r: 16 500, 15 500 and 13 500) were identified in the pAF preparation. The pAF was precipitable by Ca⁺⁺ and did not cross-react with antisera against homologous purified secondary aggregation factor and lectin. It is mainly composed of protein (48.0%) and carbohydrate (50.2%). The isolated pAF restored the aggregation potency not only of factor-depleted Geodia cells but also of cells from other Demospongiae. However, the pAF displayed no aggregation enhancing effect on urea-treated cells from species belonging to the Calcispongiae or Hexactinellida. We hypothesize that in contrast to the secondary aggregation, the initial aggregation of Geodia cells is mediated by the one-component system, the bivalent and bifunctional pAF.     

Reporter molecules as probes of DNA conformation: structure of a crystalline complex containing 2-methyl-4-nitroaniline ethylene dimethylammonium hydrobromide--5-iodocytidylyl (3'-5')guanosine

2-Methyl-4-nitroaniline ethylene dimethylammonium hydrobromide forms a crystalline complex with the self-complementary dinucleoside monophosphate, 5- iodocytidylyl (3'-5')guanosine. The crystals are tetragonal, with a = b = 32.192 A and c = 23.964 A, space group P4(3)2(1)2. The structure has been solved to atomic resolution by Patterson and Fourier methods, and refined by full matrix least squares. 5- Iodocytidylyl (3'-5')guanosine molecules are held together in pairs through Watson-Crick base-pairing, forming an antiparallel duplex structure. Nitroaniline molecules stack above and below guanine-cytosine pairs in this duplex structure. In addition, a third nitroaniline molecule stacks on one of the other two nitroaniline molecules. The asymmetric unit contains two 5- iodocytidylyl (3'-5')guanosine molecules, three nitroaniline molecules, one bromide ion and thirty-one water molecules, a total of 160 atoms. Details of the structure are described.     

Prevention of aortic lesions and hyperlipidaemia by alfalfa seed extract in cholesterol fed rabbit

Extract of alfalfa seed (ethanolic 50 % v/v) prevents the development of plaque formation and hyperlipidaemia in cholesterol fed rabbits. It inhibits the elevation of serum total cholesterol, triglycerides, phospholipids, LDL-cholesterol and total cholesterol/phospholipid ratio, while HDL-cholesterol/total cholesterol ratio increases, which is associated with a reduced incidence of atherosclerosis. Further reduction in total cholesterol and phospholipid contents of liver and heart muscle are suggestive of a beneficial role of the seed extract. The possible mechanisms of action are discussed.     

Isolation of auxotrophic mutants in support of ammonia assimilation via glutamine synthetase in Methanobacterium ivanovii

Various enzymes involved in the initial metabolic pathway for ammonia assimilation by Methanobacterium ivanovii were examined. M. ivanovii showed significant activity of glutamine synthetase (GS). Glutamate synthase (GOGAT) and alanine dehydrogenase (ADH) were present, wheras, glutamate dehydrogenase (GDH) was not detected. When M. ivanovii was grown with different levels of NH ₊ ⁴ (i.e. 2, 20 or 200 mM), GS, GOGAT and ADH activities varied in response to NH ₊ ⁴ concentration. ADH was not detected at 2 mM level, but its activity increased with increased levels of NH ₊ ⁴ in the medium. Both GS and GOGAT activities increased with decreasing concentrations of NH ₊ ⁴ and were maximum when ammonia was limiting, suggesting that at low NH ₊ ⁴ levels, GS and GOGAT are responsible for ammonia assimilation and at higher NH ₊ ⁴ levels, ADH might play a role. Metabolic mutants of M. ivanovii that were auxotrophic for glutamine were obtained and analyzed for GS activity. Results indicate two categories of mutants: i) GS-deficient auxotrophic mutants and ii) GS-impaired auxotrophic mutants.Abbreviations GS Glutamine synthetase - GOGAT glutamate synthase - GDH glutamate dehydrogenase - ADH alanine dehydrogenase     

Population biology of Avena

Summary Surveys for polymorphisms in natural populations of A. barbata sampled in California grasslands had provided evidence for widespread monomorphism and rather localized polymorphic areas in the north coastal and San Francisco regions, based on a set of morphological and isoenzymatic marker loci. Since this species, like many other annuals, was introduced from the Mediterranean region during the Spanish mission period, a comparative study of the Canadian-Welsh collections of Avena species from the Mediterranean region was undertaken using various plant characters and starch gel electrophoresis to analyze variants for esterase, phosphatase and peroxidase systems. A total of 96 samples including 73 of A. barbata and 23 of A. hirtula were studied and the results were scored to compute the polymorphism indices. In both species, only 10 to 15 percent sites showed any significant degree of polymorphism of which a majority seemed to originate from localized regions in Italy and Turkey; a part of this observed lack of within-sample variation might be the result of small sample size. In general, the patterns of variation in A. barbata from the California surveys and the present analyses seemed to be very similar and raised some interesting questions on (a) the colonizing history of introduced materials (b) the factors underlying such marked patterns of geographical variation, and (c) the current evolutionary changes occurring in these two broad, disjunct areas of species distribution.

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Zusammenfassung Untersuchungen der Polymorphismen in natürlichen Populationen von A. barbata im kalifornischen Weideland hatten einerseits zum Nachweis eines weit verbreiteten Monomorphismus und andererseits streng lokalisierter polymorpher Bereiche in der nördlichen Küsten- und San Francisco-Region geführt, wobei eine Anzahl morphologischer und isoenzymatischer Markerloci zugrunde gelegt wurde. Da diese Art, wie viele andere Annuelle auch, während der spanischen Missionsperiode aus der Mittelmeerregion eingeführt wurde, wurde eine vergleichende Untersuchung der Canadian-Welsh-Sammlungen von Avena-Arten aus der Mittelmeerregion anhand verschiedener Merkmale der Pflanzen und der Stärkegelelektrophorese-Untersuchung auf Esterase-, Phosphatase- und Peroxydase-Systeme durchgeführt. Es wurde eine Gesamtheit von 96 Stichproben, bestehend aus 73 A. barbata und 23 A. hirtula, untersucht und die Ergebnisse zur Berechnung von Polymorphismus-Indices verwendet. In beiden Arten zeigten nur 10 bis 15% der Herkünfte einen signifikanten Polymorphismusgrad. Von ihnen scheint die Mehrzahl von lokalisierten Regionen in Italien und Griechenland abzustammen. Ein Teil des beobachteten Fehlens einer Variation innerhalb der Stichproben könnte eine Folge des geringen Stichprobenumfangs sein. Im allgemeinen scheinen die Variationsmuster der kalifornischen Untersuchungen und die der vorliegenden Analysen von A. barbata sehr ähnlich zu sein. Das führt zu einigen interessanten Fragen nach a) der Besiedelungsgeschichte des eingeführten Materials, b) den Faktoren, die derart auffallenden Mustern der geographischen Variation unterliegen und c) den laufenden evolutionären Änderungen, die in diesen beiden großen, voneinander getrennten Gebieten der Artverteilung auftreten.

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This work was supported in part by a grant from the National Science Foundation (GB 8627).