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91.
92.
N-Glycoloylneuraminic acid (Neu5Gc) is synthesized as its CMP-giycosideby the action of CMPN-acetylneuramlnic acid (CMP-Neu5Ac) hydroxylase.This enzyme is a soluble cytochrome bs-dependent monooxygenaseand has been purified to apparent homogeneity from pig submandibularglands by precipitation with N-cetyN,N,N-trimethylam-moniumbromide and fractionation on Q-Sepharose, Cibacron Blue 3GA-Agarose,Reactive Brown 10-Agarose, Hexyl-Agarose and Superose S.12.This procedure resulted in an 8960-fold purification of thehydroxylase with a recovery of 0.8%. The molecular mass of thisprotein was shown to be 65 kDa on SDS-PAGE and 60 kDa as determinedby gel filtration on Superose S.12, which suggests that theenzyme is a monomer. The purified CMP-Neu5Ac hydroxylase isactivated by FeSO4 and inhibited by iron-binding reagents suchas o-phenanthroline, KCN, Tiron and ferro-zine. An apparentKm of 11 µM was determined for the substrate CMP-Neu5Acusing purified hydroxylase in the presence of Triton X-100-solubilizedmicrosomes. In a reconstituted system consisting of purifiedhydroxylase, cytochrome b5, cytochrome b5 reductase and catalase,an apparent Km of 3 µM was measured. The apparent Kmforcytochrome b5 in this system was 0.24 µM. Immunizationof a rabbit with enriched and purified hydroxylase led to anantiserum that inhibited CMP-Neu5Ac hydroxylase activity andreacted with the purified 65 kDa protein on a Western blot afterSDS-PAGE. Antibodies specific for this 65 kDa protein were isolatedand showed a strong reaction with the purified CMP-Neu5Ac hydroxylasefrom mouse liver after immunoblotting. Initial experiments withthis monospecific antibody suggest that the activity of thehydroxylase in a particular tissue correlates with the amountof immuno-reactive protein. cytochrome b5 N-glcoloylneuraminic acid hydroxylase pig submandibular gland mucin sialic acid 相似文献
93.
Modification of sialic acids by 9-O-acetylation is detected in human leucocytes using the lectin property of influenza C virus 总被引:1,自引:0,他引:1
Zimmer Gert; Suguri Toshiaki; Reuter Gerd; Yu Robert K.; Schauer Roland; Herrler Georg 《Glycobiology》1994,4(3):343-349
Influenza C virus spike glycoprotein HEF specifically recognizesglycoconjugates containing 9-O-acetyl-N-acetylneuraminic acid.The same protein also contains an esterase activity. Takingadvantage of these two properties, influenza C virus was usedas a very sensitive probe for the detection of traces of 9-O-acetyl-N-acetylneuraminicacid in human leucocytes. The binding of influenza C virus toleucocyte glycoproteins and gangliosides separated by sodiumdodecyl sulphatepolyacrylamide gel electrophoresis andthin-layer chromatography, respectively, was assayed using achromogenic esterase substrate. In this way, glycoproteins ofB-lymphocytes and T-lymphocytes were found to contain 9-O-acetylatedsialic acids. Of the various 9-O-acetylated gangliosides detected,one had the characteristics of 9-O-acetylated GD3. The identificationof 9-O-acetylated sialic acids on distinct glycoproteins andglycolipids should be helpful in assigning a physiological roleto this sugar. O-acetylation gangliosides influenza C virus lymphocytes sialic acids 相似文献
94.
Maryvonne Le Coniat Elisabeth Traiffort Martial Ruat Jean-Michel Arrang Roland Berger 《Human genetics》1994,94(2):186-188
We have assigned the human histamine H1-receptor gene to chromosome 3 by Southern blot analysis of a chromosome mapping panel constructed from humanhamster somatic cell hybrids. This assignment was confirmed by in situ hybridization on metaphase chromosomes and involved bands 3p14–p21. 相似文献
95.
Cloning and nucleotide sequence analysis of the Lactobacillus delbrueckii ssp. lactis DSM7290 cysteine aminopeptidase gene pepC 总被引:1,自引:0,他引:1
Abstract A genomic library of Lactobacillus delbrueckii ssp. lactis DSM7290 in the low copy number vector pLG339, was screened for the presence of peptidase genes. Using the chromogenic substrate gly-ala-β-naphthylamide, which is not a substrate for any of the recently cloned peptidases of DSM7290, and the multiple peptidase deficient Escherichia coli strain CM89, allowed the isolation of clones, which contained the equivalent hydrolytic activity. To identify genes encoding the conserved catalytic active site of cysteine proteases, partial nucleotide sequencing with a degenerate oligonucleotide was performed on recombinant plasmids isolated from such clones. This allowed to identify two out of nine clones to carry the Lactobacillus pepC gene. A total of 2026 nucleotides were determined, and sequence analysis revealed a gene with strong homology to the recently cloned Lb. helveticus (73.2%) and Lactococcus lactis (51.03%) pepC genes, and the derived protein showed homology with the active site of a large number of cysteine proteases. The predicted open reading frame consists of 449 codons, coding for a protein of 50 909 Da. The enzyme is functional and extremely overexpressed in E. coli . 相似文献
96.
Yong Soo Park Zhong Ping Shi Sumihisa Shiba Cayuela Chantal Shinji Iijima Takeshi Kobayashi 《Applied microbiology and biotechnology》1993,38(5):649-655
Fuzzy reasoning was applied to control both ethanol and glucose concentrations in fed-batch cultures of baker's yeast. This fuzzy controller consisted of three membership functions (concentrations of dissolved oxygen (DO), ethanol and glucose) and 18 production rules. Fuzzy inference was carried out by IF {A is a and B is b,...#x007D;, THEN {C is c} from the on-line measured concentrations of DO, ethanol and glucose. When medium concentrations of ethanol and glucose in fed-batch culture of baker's yeast were set at 2 g/l and 0.2 g/l, both ethanol and glucose concentrations were controlled at 2.67±0.35 g/l and 0.27±0.25 g/l, respectively, ethanol production was reduced from 26 g/l to 34 g/l, cell yield increased from 0.38 to 0.53 g dry cell/g consumed glucose and ethanol yield decreased from 0.50 to 0.14 g ethanol/g consumed glucose, respectively, as compared with those of the glucose only control at 0.2 g/l. 相似文献
97.
The variation pattern in the perennial Hordeum brachyantherum complex and in the annual H. depressum are described. The diploid form of H. brachyantherum s. lat., endemic to California in USA, previously recognized as a separate species is here treated as a subspecies ( H. brachyantherum ssp. californicum ). Despite its restricted distribution it shows a considerable variation and overlap in morphology with the tetraploid ssp. brachyantherum , and no unambiguous determination based on morphology between the two tax a is possible. The tetraploid cytotype has a large distribution area in western North America and easternmost Asia and a very wide morphological variation. It has also a small disjunct distribution area in easternmost Canada. A single hexaploid population from California is referred to ssp. brachyantherum . 相似文献
98.
Per Mrin Roland Rosmond Bengt-ke Bengtsson Carola Gustafsson Gran Holm Per Bjmtorp 《Obesity (Silver Spring, Md.)》1994,2(3):263-270
Visceral obese men were characterized by a decreased total GH secretion and diminished peak amplitude, size, and number. T-substitution was followed by elevation of IGF-I levels. The IGF-I increase correlated with the elevation of T-concentration, and was most pronounced in men with the lowest concentrations of free T from the outset. There were no detectable changes in total quantity, amplitude, size or number of peaks of GH secretion. Glucose, cholesterol and triglycerides as well as diastolic blood pressure decreased. There were no changes in thyroid or hematology variables. Visceral obesity in men has been reported to be characterized by low testosterone (T) and insulin-like growth factor I (IGF-I) concentrations, the latter suggesting a relative growth hormone (GH) deficiency. Since T and GH-secretions are interrelated, men with visceral obesity were substituted with T for 14 days, and diurnal secretion pattern of GH as well as IGF-I concentrations, and metabolic variables were followed. T-substitution of visceral obese men is followed by an elevation of IGF-I concentrations. It is suggested that this might be due either to minor, non-detectable increases in GH secretion, or to direct effects of T on IGF-I concentrations. The regulatory mechanisms by which T-administration are leading to metabolic and anthropometric improvements, might be direct effects of T, with or without mediation via GH secretion. 相似文献
99.
The influence of catchment size on lake trophic status during the hemlock decline and recovery (4800 to 3500 BP) in southern Ontario lakes 总被引:3,自引:1,他引:2
We investigated the ecological effects of terrestrial ecosystem change during the hemlock decline and recovery (4,800–3,500 BP) on lake communities (diatoms and chrysophytes). This study specifically assessed the role of catchment area and slope in determining the magnitude of lake eutrophication during the hemlock decline by analyzing sediment cores from five alkaline, holomictic lakes in southeastern Ontario, Canada. The study lakes were similar in most limnological aspects, but differed widely in the relative sizes of their catchments. Diatoms were used to quantitatively infer past lake-water total phosphorus (TP) concentrations.All five lakes showed shifts in their algal communities during the hemlock decline, but most lakes exhibited only minor changes in trophic status. The magnitude of the limnological response appears to be related to catchment size and slope. Long Lake, Burridge Lake, and Gunter Lake possess the smallest catchments and exhibited the weakest responses to the hemlock decline. The catchment area of Flower Round Lake is considerably larger and steeper than these lakes, and was the only lake to show a marked eutrophication. Aulacoseira ambigua bloomed and diatom-inferred TP concentration increased by 14 µg 1–1.Catchment slope appears to have influenced the type of material exported into the lakes. Lake basins draining catchments with gentle relief received proportionally greater amounts of organic matter, whereas steeper catchments supplied relatively greater proportions of mineral matter. Faster water flow associated with steeper catchment slope may have enhanced mineral erosionFollowing the hemlock decline, nutrient supplies to most of the study lakes were reduced. The period of forest recovery was associated with an 11 µg 1–1 reduction in diatom-inferred lake-water TP concentration in Flower Round Lake, and algal populations decreased. Our results generally support the ecological theory of forest ecosystem development and secondary succession developed from long-term data collected at the Hubbard Brook Experimental Ecosystem. 相似文献
100.
Roland Rost 《Journal of Ornithology》1993,134(1):93-95
Summary A pair of Marsh Tits ( white/blue; yellow/green), which has been caught for bioacoustical investigations in May 1984, was released at the same place 15 months later (August 1985). Although their territory had been occupied by another Marsh Tit pair, the released birds reconquered it after violent skirmishes within one day. In 1986 White/Blue and Yellow/Green raised successfully eight young. Because its female partner died probably during winter 1986/87, in 1987, 1988 and 1989 Yellow/Green was observed raising young together with a female (Blue) ringed in autumn 1986 as a this year's bird. On the 15th of January 1990 I saw Yellow/Green (at least seven years and eight months old) for the last time. Although White/Blue and Yellow/Green were kept in captivity for 15 months, they obviously were able to maintain themselves free living after they had been released again. Thus, small passerines — when housed in accordance with their species specific claims and handled properly — are definitely able to survive in the wild even if they had lived in captivity for longer periods. 相似文献