scholarly journals Aberrant 3H labelling of ATP during in vivo labelling of Ehrlich mouse ascites tumour cells with [2-3H]inositol is significant in the study of isomers of InsP3 and InsP4

1994 ◽  
Vol 300 (3) ◽  
pp. 859-863 ◽  
Author(s):  
S Christensen ◽  
H Harbak ◽  
L O Simonsen
Keyword(s):  
Anion Exchange ◽  
Ambient Pressure ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Tumour Cells ◽  
Ascites Tumour ◽  
Small Column ◽  
Mouse Ascites ◽  
Ascites Tumour Cells

Neutralized perchloric acid extracts of intra-abdominally proliferating Ehrlich mouse ascites-tumour cells harvested after 24 h exposure to [2-3H]inositol were analysed by Mono Q HR5/5 anion-exchange h.p.l.c. using an ammonium formate/phosphoric acid gradient or by ambient-pressure small-column anion-exchange chromatography (Bio-Rad AG 1-X8, 200-400 mesh). The results showed that cellular ATP contained aberrant 3H label in excess of 3H in the isomers of InsP3 and InsP4. The putative ATP 3H radioactivity showed: (i) h.p.l.c. run time as the material causing the largest A254 peak traced, (ii) precise spiking with ATP and [14C]ATP and (iii) specific absorption to charcoal. Moreover, enzymic conversion of ATP into ADP changed putative ATP 3H into putative ADP 3H. In addition, aberrant 3H labelling of cellular ADP and GTP was detected, although at a lower level.

Purification and characterization of a Ca2+-dependent novel lectin from Nymphaea nouchali tuber with antiproliferative activities

2011 ◽  
Vol 31 (6) ◽  
pp. 465-475 ◽  
Author(s):  
Syed Rashel Kabir ◽  
Md. Abu Zubair ◽  
Md. Nurujjaman ◽  
Md. Azizul Haque ◽  
Imtiaj Hasan ◽  
...  
Keyword(s):  
Anion Exchange ◽  
Pathogenic Bacteria ◽  
Sequence Similarity ◽  
Deae Cellulose ◽  
Ehrlich Ascites Carcinoma ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Neutral Sugar ◽  
The Stability

A lectin (termed NNTL) was purified from the extracts of Nymphaea nouchali tuber followed by anion-exchange chromatography on DEAE-cellulose, hydrophobic chromatography on HiTrap Phenyl HP and by repeated anion-exchange chromatography on HiTrap Q FF column. The molecular mass of the purified lectin was 27.0 ± 1.0 kDa, as estimated by SDS/PAGE both in the presence and in the absence of 2-mercaptoethanol. NNTL was an o-nitrophenyl β-D-galactopyranoside sugar-specific lectin that agglutinated rat, chicken and different groups of human blood cells and exhibited high agglutination activity over the pH range 5–9 and temperatures of 30–60°C. The N-terminal sequence of NNTL did not show sequence similarity with any other lectin and the amino acid analysis revealed that NNTL was rich in leucine, methionine and glycine residues. NNTL was a glycoprotein containing 8% neutral sugar and showed toxicity against brine shrimp nauplii with an LC50 value of 120 ± 29 μg/ml and exerted strong agglutination activity against four pathogenic bacteria (Bacillus subtilis, Sarcina lutea, Shigella shiga and Shigella sonnei). In addition, antiproliferative activity of this lectin against EAC (Ehrlich ascites carcinoma) cells showed 56% and 76% inhibition in vivo in mice at 1.5 and 3 mg·kg−1·day−1 respectively. NNTL was a divalent ion-dependent glycoprotein, which lost its activity markedly in the presence of denaturants. Furthermore, measurement of fluorescence spectra in the presence and absence of urea and CaCl2 indicated the requirement of Ca2+ for the stability of NNTL.

scholarly journals Ionophore-mediated coupling between ion fluxes and amino acid absorption in mouse ascites-tumour cells. Restoration of the physiological gradients of methionine by valinomycin in the absence of adenosine triphosphate

1974 ◽  
Vol 140 (3) ◽  
pp. 383-393 ◽  
Author(s):  
M. Reid ◽  
L. E. Gibb ◽  
A. A. Eddy
Keyword(s):  
Amino Acid ◽  
Sodium Pump ◽  
Tumour Cells ◽  
Ascites Tumour ◽  
Maximum Value ◽  
Methionine Concentration ◽  
Mouse Ascites ◽  
Na Ions ◽  
Physiological Gradients ◽  
Ascites Tumour Cells

1. Preparations of mouse ascites-tumour cells depleted of ATP and Na+ ions accumulated l-methionine, in the presence of cyanide and deoxyglucose, from a 1mm solution containing 80mequiv. of Na+/l and about 5mequiv. of K+/l. Valinomycin increased, from about 4 to 16, the maximum value of the ratio of the cellular to extracellular concentrations of methionine formed under these conditions without markedly affecting the distributions of Na+ and of K+. Similar observations were made with 2-aminoisobutyrate, glycine and l-leucine. Increasing the extracellular concentration of K+ progressively decreased the accumulation of methionine in the presence of valinomycin. Over the physiological range of ionic gradients, the system behaved as though the absorption of methionine with Na+ was closely coupled to the electrogenic efflux of K+ through the ionophore. The process was insensitive to ouabain and so the sodium pump was probably not involved. 2. The amount of methionine accumulated during energy metabolism was similar to the optimal accumulation in the presence of valinomycin when ATP was lacking. It was also similarly affected by increasing the methionine concentration. 3. A mixture of nigericin and tetrachlorosalicylanilide mimicked the action of valinomycin. The anilide derivative inhibited the absorption of 2-aminoisobutyrate in the presence of valinomycin, but not in its absence. 4. Gramicidin inhibited methionine absorption and caused the preparations to absorb Na+ and lose K+. 5. The observations appear to verify the principle underlying the gradient hypothesis by showing that the tumour cells can efficiently couple the electrochemical gradient of Na+ to the amino acid gradient.

scholarly journals Metabolism and sodium transfer of mouse ascites tumour cells

1958 ◽  
Vol 140 (1) ◽  
pp. 80-93 ◽  
Author(s):  
M. Maizels ◽  
Mary Remington ◽  
R. Truscoe
Keyword(s):  
Tumour Cells ◽  
Ascites Tumour ◽  
Sodium Transfer ◽  
Mouse Ascites ◽  
Ascites Tumour Cells

scholarly journals Characterisation of a cell swelling‐activated K + ‐selective conductance of Ehrlich mouse ascites tumour cells

2000 ◽  
Vol 524 (3) ◽  
pp. 757-767 ◽  
Author(s):  
María Isabel Niemeyer ◽  
Charlotte Hougaard ◽  
Else K. Hoffmann ◽  
Finn Jørgensen ◽  
Andrés Stutzin ◽  
...  
Keyword(s):  
Tumour Cells ◽  
Cell Swelling ◽  
Ascites Tumour ◽  
A Cell ◽  
Mouse Ascites ◽  
Ascites Tumour Cells
Sign in / Sign up

Export Citation Format

Share Document