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1.
Chromatin was prepared from the buds and cotyledons of Alaskapea seedlings. The dissociated chromosomal components in thepresence of 2 M NaCl and 5 M urea were completely fractionatedinto DNA and proteins with a Bio-Gel A50 column. The proteinswere recovered by (NH4)2SO4 and further fractionated into histonesand non-histone proteins using a Bio-Rex 70(Na+) column. Thedifference in the ratios of histones to non-histone proteinsbefore and after chromatography with the Bio-Rex 70 was lessthan 10%. The histones and non-histone proteins thus preparedshowed typical protein absorption spectra. Polyacrylamide gelelectrophoresis of histones showed that the histone compositionsin buds and cotyledon were similar, but the amount of HI histoneswas a little less in cotyledons than in buds. Unlike histones,non-histone proteins fractionated by SDS-polyacrylamide gelelectrophoresis indicated distinct differences between the twotissues. Buds had more heterogeneous non-histone proteins, atleast 13 polypeptides, than cotyledons did. On the other hand,non-histone proteins of cotyledons showed less heterogeneityand lacked proteins of high molecular weight which were foundin buds. (Received May 6, 1976; ) 相似文献
2.
Eiichi Tachikawa A. William Tank David H. Weiner Werner F. Mosimann Nobuyuki Yanagihara Norman Weiner 《Journal of neurochemistry》1987,48(5):1366-1376
Abstract: Incubation of rat pheochromocytoma PC12 cells with 4β-phorbol-12β-myristate-13α-acetate (PMA), an activator of Ca2+/phospholipid-dependent protein kinase (protein kinase C), or forskolin, an activator of adenylate cyclase, is associated with increased activity and enhanced phosphorylation of tyrosine hydroxylase. Neither the activation nor increased phosphorylation of tyrosine hydroxylase produced by PMA is dependent on extracellular Ca2+. Both activation and phosphorylation of the enzyme by PMA are inhibited by pretreatment of the cells with trifluo-perazine (TFP). Treatment of PC 12 cells with l-oleoyl-2-acetylglycerol also leads to increases in the phosphorylation and enzymatic activity of tyrosine hydroxylase; 1, 2-diolein and 1, 3-diolein are ineffective. The effects of forskolin on the activation and phosphorylation of the enzyme are independent of Ca2+ and are not inhibited by TIT5. Forskolin elicits an increase in cyclic AMP levels in PC 12 cells. The increases in both cyclic AMP content and the enzymatic activity and phosphorylation of tyrosine hydroxylase following exposure of PC 12 cells to different concentrations of forskolin are closely correlated. In contrast, cyclic AMP levels do not increase in cells treated with PMA. Tryptic digestion of the phosphorylated enzyme isolated from untreated cells yields four phosphopeptides separable by HPLC. Incubation of the cells in the presence of the Ca2+ ionophore ionomycin increases the phosphorylation of three of these tryptic peptides. However, in cells treated with either PMA or forskolin, there is an increase in the phosphorylation of only one of these peptides derived from tyrosine hydroxylase. The peptide phosphorylated in PMA-treated cells is different from that phosphorylated in forskolin-treated cells. The latter peptide is identical to the peptide phosphorylated in dibutyryl cyclic AMP-treated cells. These results indicate that tyrosine hydroxylase is activated and phosphorylated on different sites in PC 12 cells exposed to PMA and forskolin and that phosphorylation of either of these sites is associated with activation of tyrosine hydroxylase. The results further suggest that cyclic AMP-dependent and Ca2+/ phospholipid-dependent protein kinases may play a role in the regulation of tyrosine hydroxylase in PC 12 cells. 相似文献
3.
Photosynthetically competent chloroplasts were isolated fromcells of Euglena gracilis Z grown photoautotrophically in 1.5%CO2. The isolated chloroplasts were intact and substantiallyfree from cytosolic, mitochondrial and microbody materials.The effects of some compounds on the activity of photosynthetic14CO2 fixation were examined. The optimal pH and sorbitol concentrationwere 8.0 and 0.33 M, respectively. The chloroplasts requireda high level of P, (5 to 20 mM) for the maximal rate of photosynthesis.They were insusceptible to 10 mM of free Mg2+. ATP, ADP andAMP at 1 to 5 mM notably stimulated photosynthesis, althoughhigh concentrations of AMP were unfavorable. In the assay mediumdeveloped for this study, the chloroplasts exhibited photosyntheticactivity of 120µmoles-mg1 Chl-h1 at 30?C. Chloroplasts could also be isolated from cells grown under ordinaryair. The rate of photosynthetic 14CO2 fixation at 1 mM NaHl4CO3was higher in these chloroplasts than in those isolated fromcells grown in 1.5% CO2, whereas at 10 mM NaHl4CO3, the ratesof the two types of chloroplasts were nearly the same. Theseresults suggest that the CO2 concentration given during growthof the algal cells affects the affinity for dissolved inorganiccarbon at the chloroplast level. (Received March 30, 1987; Accepted August 17, 1987) 相似文献
4.
In cells of cyanobacterium Anabaena variabilis grown under ordinaryair (low-CO2 cells), the transport of both CO2 and HCO3was significantly enhanced by Na+. This effect was pronouncedas the external pH increased. When low-CO2 cells were treatedwith an inhibitor of carbonic anhydrase (CA), only CO2 transportbut not HCO3 transport, was inhibited. The initial rateof photosynthetic carbon fixation as a function of the concentrationof internal inorganic carbon (IC) was practically the same irrespectiveof whether CO2 or HCO3 was externally supplied. Theseresults suggest that IC is actively transported through theplasma membrane in a form of HCO3 probably by some transporterand that the transmembrane Na+ gradient is involved in thisIC transport system. Free CO2 may be hydrated by CA to HCO3and then transported to the cells by this transporter. On the other hand, CO2 is actively taken up by cells grown withair containing 5% CO2 (high-CO2 cells) though the enhancingeffect of Na+ was much smaller in high- CO2 cells than in low-CO2cells. The initial rate of fixation as a function of internal IC concentrationindicated that the rate of the carboxylation reaction of accumulatedIC is higher in I0W-CO2 cells than in high-CO2 cells. The studieswith ethoxyzolamide indicated that even in low-CO2 cells, CAdoes not function inside Anabaena cells. These results suggestthat inside the low-CO2 cells of Anabaena, some mediator(s)facilitates the transport of IC to RuBPCase. (Received January 23, 1987; Accepted April 24, 1987) 相似文献
5.
Inorganic carbon transport during photosynthesis of cyanobacteriumAnabaena variabilis grown under ordinary air was investigatedby supplying 14CO2 or H14CO3 solution to three differentstrains. Both CO2 and HCO3 were accumulated within thealgal cells. In the cell suspension from which dissolved inorganiccarbon had been depleted by pre-illumination, CO2 was transportedand accumulated faster than HCO3. When the concentrationof HCO3 injected into the cell suspension of A. variabilisM3 was 25 times as high as that of CO2 (the expected ratio atequilibrium at pH 7.8), the initial rates of fixation of bothinorganic carbon species were practically the same. On the otherhand, when 14CO2 or H14CO3 was added under steady statephotosynthetic conditions, both carbon species were transportedat similar rates. The ratio of fixed to transported carbon measuredafter the initial 5 s was only 2327% regardless of thecarbon species supplied. This percentage is much lower thanthat reported for Chlorella cells.
1 To whom reprint requests should be addressed (Received June 30, 1986; Accepted December 16, 1986) 相似文献
6.
Acidic inorganic phosphate (Pi) pool (pH around 6) was detected besides the cytoplasmic pool in intact cells of Chlorella vulgaris 11h by 31P-in vivo nuclear magnetic resonance (NMR) spectroscopy. It was characterized as acidic compartments (vacuoles) in combination with the cytochemical technique; staining the cells with neutral red and chloroquine which are known as basic reagents specifically accumulated in acidic compartments. Under various conditions, the results obtained with the cytochemical methods were well correlated with those obtained from in vivo NMR spectra; the vacuoles were well developed in the cells at the stationary growth phase where the acidic Pi signal was detected. In contrast, cells at the logarithmic phase in which no acidic Pi signal was detected contained only smaller vesicles that accumulated these basic reagents. No acidic compartment was detected by both cytochemical technique and 31P-NMR spectroscopy when the cells were treated with NH4OH. The vacuolar pH was lowered by the anaerobic treatment of the cells in the presence of glucose, while it was not affected by the external pH during the preincubation ranging from 3 to 10. Possible vacuolar functions in unicellular algae especially with respect to intracellular pH regulation are discussed.Non-standard abbreviations EDTA
ethylenediaminetetraacetic acid
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid
- MDP
methylene diphosphonic acid
- NMR
nuelear magnetic resonance
- PCA
perchloric acid
- PCV
packed cell volume
- Pi
inorganic phosphate
- Pic
sytoplasmic inorganic phosphate
- Piv
vacuolar inorganic phosphate
- ppm
parts per million
- SP
sugar phosphates
- TCA
trichloroacetic acid 相似文献
7.
Masanori Ito Kazuhiro Yoshida Eikai Kyo Ayse Ayhan Hirofumi Nakayama Wataru Yasui Hisao Ito Eiichi Tahara 《Virchows Archiv. B, Cell pathology including molecular pathology》1990,59(1):173-178
We have examined the expression of mRNAs for epidermal growth factor (EGF), transforming growth factor-alpha (TGF-α), EGF
receptor (EGFR), PDGF-A chain (PDGFA), PDGF-B chain (PDGFB) and PDGF receptor (PDGFR) genes in seven human colorectal carcinoma
cell lines and 18 human colorectal carcinomas.
In surgically resected specimens of the 18 colorectal tumors, TGF-α, EGFR, PDGFA, PDGFB and PDGFR mRNAs were detected at various
levels in 15 (83%), 9 (50%), 18 (100%), 8 (44%) and 12 (67%), respectively. They were also detected in normal tissues. Interestingly,
EGF mRNA was detected in only five (28%) of the tumors, but not in normal mucosa. Expression of EGF was also confirmed immunohistochemically
in tumor cells. Of the five tumors expressing EGF, four expressed EGFR mRNA and showed a tendency to invade veins and lymphatics.
All the colorectal carcinoma cell lines expressed TGF-α mRNA, and five cell lines expressed EGFR mRNA simultaneously. Production
of TGF-α protein by DLD-1 and CoLo320DM cells was confirmed by TGF-α specific monoclonal antibody binding assay. The spontaneous3H-thymidine uptake by DLD-1 was suppressed by an anti-TGF-α monoclonal antibody. PDGFA and PDGFB mRNA were also expressed
in four cell lines, but PDGFR and EGF mRNA was not detected. These results suggest that human colorectal carcinomas express
multi-loops of growth factors and that TGF-α produced by tumor cells functions as an autocrine growth factor in human colonic
carcinoma. 相似文献
8.
Salt tolerance of the reed plant Phragmites communis 总被引:6,自引:0,他引:6
Reed plants ( Phragmites communis Trinius) were grown at NaCl concentrations up to 500 m M and their growth, mineral contents and leaf blade osmotic potential were determined. Addition of NaCl up to 300 m M did not affect growth significantly. Sucrose, Cl- and Na+ concentrations in the shoots increased with the salinity of the medium and the shoot water content decreased. K+ always contributed most to the leaf osmotic potential. Even in the presence of 250 m M NaCl in the rooting medium, the leaf blade contained only 50 mM Na+ , suggesting that the plants have an efficient mechanism for Na+ exclusion. 22 Na+ uptake experiments suggested that the retranslo-cation of absorbed Na+ from shoots to the rooting medium lowered the uptake of Na+ . 相似文献
9.
Root elongation of Alaska pea seedling was suppressed by higherconcentrations of growth retardants, CCC and ancymidol, thanthose required for shoot elongation. Gibberellic acid (GA3)led to recovery of ancymidol-inhibited elongation, with theconcentration (1 nM) required for roots being lower than thatfor shoots (10 µM). Ancymidol caused swelling of corticalcells in the elongating zone of the root, while GA3 completelycanceled this. These results suggest that roots require muchless gibberellin than shoots for normal elongation growth. Growth kinetics recorded by a computer-regulated rhizometerindicated that the lag periods for growth suppression by ancymidoland growth recovery by GA3 were about 10 h and 7 h, respectively. The composition of the cell wall sugars changed remarkably alongthe root axis from the tip to the base. The arabinose contentwas highest in the tip and rapidly decreased toward the base,whereas galactose complementarily increased toward the base.The thickened zone of ancymidol-treated roots had a higher galactosecontent than GA3-treated slender roots. Other neutral sugarswere not significantly influenced by ancymidol and/or GA3. Theseresults suggest that ancymidol makes cells short and thick withgalactose-rich cell walls while GA3 keeps cells extensible andslender with galactose-poor cell walls. (Received March 3, 1987; Accepted December 4, 1987) 相似文献
10.
Summary The pyrenoid is a protein complex in the chloroplast stroma of eukaryotic algae. After the treatment with mercury chloride, pyrenoids were isolated by sucrose density gradient centrifugation from cell-wall less mutant cells, CW-15, as well as wild type cells, C-9, of unicellular green algaChlamydomonas reinhardtii. Pyrenoids were characterized as a fraction whose protein/chlorophyll ratio was very high, and also examined by Nomarski differential interference microscopy. Most of the components consisted of 55 kDa and 16 kDa polypeptides (11) which were immunologically identified as the large and small subunit of RuBisCO (ribulose-1,5-bisphosphate carboxylase/oxygenase) protein, respectively. Some minor polypeptides were also detected. Substantial amount of RuBisCO protein is present as a particulate form in the pyrenoid in addition to the soluble form in algal chloroplast stroma.Abbreviations BPB
bromophenol blue
- DAB
3,3-diaminobenzidine
- DTT
dithiothreitol
- ELISA
enzyme-linked immunosorbent assay
- High-CO2 cells
cells grown under air enriched with 4% CO2
- Low-CO2 cells
cells grown under ordinary air (containing 0.04% CO2)
- NP-40
nonionic detergent (Nonidet) P-40
- PAGE
polyacrylamide gel electrophoresis
- PAP
peroxidase-antiperoxidase conjugate
- RuBisCO
ribulose-1,5-bisphosphate carboxylase/oxygenase
- RuBP
ribulose-1,5-bisphosphate
- SDS
sodium dodecylsulfate 相似文献