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91.
Characterization of Nitrate Reductase from Corn Leaves (Zea mays cv W64A x W182E) : Two Molecular Forms of the Enzyme
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The primary leaves from corn seedlings grown for 6 days were harvested, frozen with liquid N2 and extracted in a Tris buffer (pH 8.5, 250 millimolar) containing 1 millimolar dithiothreitol, 10 millimolar cysteine, 1 millimolar EDTA, 20 micromolar flavin adenine dinucleotide and 10% (v/v) glycerol. Nitrate reductase (NR) in the crude extract was stable for several days at 0°C and for several months at −80°C. The enzyme was purified using (NH4)2SO4 fractionation, brushite-hydroxyl-apatite chromatography and blue-sepharose affinity chromatography. The enzyme was eluted from the blue-sepharose column with a linear gradient of NADH (0-100 micromolar) or with 0.3 molar KNO3. About 10% of the original activity was recovered with NADH (NADH-NR). It had a specific activity of about 60 to 70 units (micromoles NO2− per minute per milligram protein). A sequential elution with NADH followed by KNO3 (0.3 molar) or KCl (0.3 molar) yielded 2 peaks. Rechromatography of each peak gave two peaks again. These results indicate that we are dealing with two forms of the same enzyme rather than two different NR proteins. The two NRs had different molecular weights as judged by chromatography on Toyopearl. The NADH-NR was more sensitive than the NO3−-NR to antibody prepared against barley leaf NR. In Ouchterlony assays a single precipitin line, with completely fused boundaries, was observed. 相似文献
92.
Sato Takahide; Kusaba Sakiku; Nakagawa Hiroki; Ogura Nagao 《Plant & cell physiology》1984,25(6):1069-1071
By using a monospecific anti-polygalacturonase-2 antibody, a54K-dalton polypeptide was detected in in vitro translationproducts by a wheat germ cell-free translation system programmedwith polyadenylated RNA from ripe tomato pericarp tissue. Thisputative precursor of polygalacturonase was about 9K daltonslarger in molecular weight than polygalacturonase-2. (Received December 12, 1983; Accepted May 17, 1984) 相似文献
93.
H Nakagawa U Izumi S Tsurufuji 《Biochemical and biophysical research communications》1977,79(1):260-266
Insoluble collagen of granulation tissue produced by carrageenin injection was solubilized by pepsin treatment and purified. The pepsin-solubilized insoluble collagen contained partially degraded collagen fragments and the amounts of these small fragments of collagen were much greater in the resorbing granulation tissues than in the growing tissues, suggesting that these small fragment were formed in the course of resorption of granulation tissue, including collagen breakdown. 相似文献
94.
Yoshihide Yamasaki Osamu Shimamura Akira Kizu Masao Nakagawa Hamao Ijichi 《Life sciences》1982,31(5):471-478
The formaldehyde method was used to examine the interaction of PGE1 with morphine, β-endorphin and Met-enkephalin on rat mast cells by their effects on IgE-mediated 14C-serotonin release. PGE1 (2×10?8?2×10?5 M) caused a dose-related inhibition of the mediator release 1 min after an antigen challenge, and morphine (3×10?7?3×10?5 M) reversed this PGE1 effect dose-dependently and stereospecifically; naloxone (2×10?4 M) antagonized this action of morphine. β-Endorphin (3×10?7?10?5 M) and Met-enkephalin (3×10?6?10?4 M) mimicked this morphine action dose-dependently and were antagonized by naloxone (2×10?4 M). These results suggest that morphine and endorphins modulate immunological mediator release from rat mast cells through opioid receptors. 相似文献
95.
Induction of proliferation and Ig production in human B leukemic cells by anti-immunoglobulins and T cell factors 总被引:20,自引:0,他引:20
K Yoshizaki T Nakagawa T Kaieda A Muraguchi Y Yamamura T Kishimoto 《Journal of immunology (Baltimore, Md. : 1950)》1982,128(3):1296-1301
The proliferation and differentiation of human leukemic B cells (B-CLL cells) with anti-Ig and T cell-derived helper factors are described. Stimulation of B-CLL cells with anti-Ig and T helper factors could induce proliferation as well as differentiation into IgM- and IgG-producing cells. Neither anti-Ig nor T helper factors alone could induce any proliferation and/or differentiation of B-CLL cells. Not only whole molecules of anti-Ig but also F(ab')2 fragments could induce proliferation and differentiation of B-CLL cells in the presence of T helper factors, but monovalent Fab' fragments were not effective. Induction of both IgM and IgG with the same idiotype was confirmed by immunofluorescent and SDS-PAGE analysis. By employing an IL 2-dependent cytotoxic T cell line and a TRF-responsive B cell line, T cell factors were separated into a fraction with IL2 activity but no TRF activity and a fraction with TRF activity but no IL 2 activity by chromatofocusing. Anti-Ig and IL 2 fraction could induce proliferation of B-CLL cells, but TRF fraction was not effective for the induction of proliferation in anti-IG-stimulated cells. For IgM and IgG production, anti-Ig and both IL 2 and TRF fractions were required. Depletion of IL 2 fraction in the first 2 days' culture inhibited Ig production, whereas the absence of TRF fraction in the first 2 days did not show any inhibitory effect on Ig production. 相似文献
96.
Pathological observation on experimental swine dysentery 总被引:1,自引:0,他引:1
M Kubo M Nakagawa M Kashiwazaki S Konno 《National Institute of Animal Health quarterly》1979,19(3):83-90
Experimental swine dysentery caused by 4 cultured strains (S73/2, DJ183, DJ70 and DK762) of Treponema hyodysenteriae was studied pathologically. The distribution and quantity of treponemes were examined on tissue sections stained by the Warthin-Starry method. Of the organs the colon contained the largest number of treponemes and the cecum and rectum the second largest number. Histopathological lesions were restricted to the large intestine. They ranged from mild catarrhal colitis in the mild case to desquamative, hemorrhagic colitis in the severest case. The severity of lesion was closely associated with the quantity of treponemes present. There was no difference in quality of the lesion between any two of the strains used in this study. Electron microscope revealed a large number of free treponemes present in the intestinal lumen and crypts. Treponemes were seen more frequently in the cytoplasm of goblet cells than in that of intestinal epithelial cells. They were also observed in desquamated degenerative epithelia. A small number of them were found in intact epithelia. Morphologically, the treponeme had a granular protoplasmic cylinder at the center which was surrounded by a thin envelope. Between the cylinder and the envelope there were axial fibrils. 相似文献
97.
A sensitivie and specific radioimmunoassay for the measurement of serum ursodeoxycholic acid has been developed. Ursodeoxycholic acid bound to bovine serum albumin was used as an antigen, and antiserum to this antigen was raised in the rabbit. [11, 12-3h2]Ursodeoxycholic acid was used as the radioactive tracer, and the radioimmunoassay was carried out by the method of Simmonds et al. (1973. Gastroenterology. 65: 705-711). The percentage of bound radioactivity decreased linearly with a logarithmic increase in unlabeled ursodeoxycholic acid from 10 to 200 pmol. The antiserum showed extremely high specificity for ursodeoxycholic acid (free and conjugated), and the values determined by radioimmunoassay indicated a close correlation with those found by gas-liquid chromatography. In normal Japanese subjects, a small amount of ursodeoxycholic acid in serum was detected, and the level was detected, and the level was 0.15 +/- 0.11 nmol/ml. This convenient radioimmunoassay will provide useful information about the metabolism of ursodeoxycholic acid in man. 相似文献
98.
The increase in the dichroism of the ester-C=O and COObands in response to mechanical extension in oat coleoptilecell walls was much enhanced by IAA pretreatment while the decreasein that of the C-O-C band was not enhanced or even suppressed.The results indicate the importance of the ultrastructure ofnon-cellulosic polysaccharides in controlling auxin-inducedcell elongation.
1 Present adress: Department of Agricultural Chemistry, KyotoPrefectural University, Shimogamo, Kyoto 606, Japan. (Received May 16, 1978; ) 相似文献
99.
Microbial Production of Ursodeoxycholic Acid from Lithocholic Acid by Fusarium equiseti M41 总被引:2,自引:2,他引:0
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Haruji Sawada Songsri Kulprecha Naline Nilubol Toshiomi Yoshida Shinichi Kinoshita Hisaharu Taguchi 《Applied microbiology》1982,44(6):1249-1252
A fungus identified as Fusarium equiseti was isolated from soil and found to carry out 7β-hydroxylation of lithocholic acid to ursodeoxycholic acid (35% yield; 350 mg/liter) in 112 h. 相似文献
100.
The conformational stability and flexibility of insulin with an additional intramolecular cross-link 总被引:1,自引:0,他引:1
D N Brems P L Brown S H Nakagawa H S Tager 《The Journal of biological chemistry》1991,266(3):1611-1615
The conformational stability and flexibility of insulin containing a cross-link between the alpha-amino group of the A-chain to the epsilon-amino group of Lys29 of the B-chain was examined. The cross-link varied in length from 2 to 12 carbon atoms. The conformational stability was determined by guanidine hydrochloride-induced equilibrium denaturation and flexibility was assessed by H2O/D2O amide exchange. The cross-link has substantial effects on both conformational stability and flexibility which depend on its length. In general, the addition of a cross-link enhances conformational stability and decreases flexibility. The optimal length for enhanced stability and decreased flexibility was the 6-carbon link. For the 6-carbon link the Gibbs free energy of unfolding was 8.0 kcal/mol compared to 4.5 kcal/mol for insulin, and the amide exchange rate decreased by at least 3-fold. A very short cross-link (i.e. the 2-carbon link) caused conformational strain that was detectable by a lack of stabilization in the Gibbs free energy of unfolding and enhancement in the amide exchange rate compared to insulin. The effect of the cross-link length on insulin hydrodynamic properties is discussed relative to previously obtained receptor binding results. 相似文献