Characterization of mononucleosomes and associated glycoproteins from Ehrlich ascites tumour cells

1980 ◽  
Vol 58 (11) ◽  
pp. 1261-1269 ◽  
Author(s):  
Brian L. A. Miki ◽  
James W. Gurd ◽  
Ian R. Brown
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Apparent Molecular Weight ◽  
Micrococcal Nuclease ◽  
Nonhistone Proteins ◽  
Ehrlich Ascites ◽  
Ehrlich Ascites Tumour ◽  
Ascites Tumour Cells ◽  
Ehrlich Ascites Tumour Cells

Mononucleosomes generated by the digestion of ascites nuclei with micrococcal nuclease were resolved into three types by polyacrylamide gel electrophoresis. All three types were present in early and late digests and a precursor–product relationship was not apparent. Each mononucleosome type was associated with a unique pattern of subnucleosome DNA fragments and differences were apparent in histone and nonhistone proteins. On the basis of reactivity to 125I-labelled concanavalin A and labelling with [3H]glucosamine, glycoproteins were identified as components of two of the mononucleosome types. The principal glycoprotein species associated with mononucleosomes had an apparent molecular weight of 130 000.

scholarly journals Purification of phosphate-dependent glutaminase from isolated mitochondria of Ehrlich ascites-tumour cells

1988 ◽  
Vol 255 (3) ◽  
pp. 1031-1035 ◽  
Author(s):  
A R Quesada ◽  
F Sanchez-Jimenez ◽  
J Perez-Rodriguez ◽  
J Marquez ◽  
M A Medina ◽  
...  
Keyword(s):  
Rat Kidney ◽  
Polyacrylamide Gel Electrophoresis ◽  
Tumour Cells ◽  
Ph Optimum ◽  
Isolated Mitochondria ◽  
Ascites Tumour ◽  
Ehrlich Ascites ◽  
Ehrlich Ascites Tumour ◽  
Ascites Tumour Cells ◽  
Ehrlich Ascites Tumour Cells

Phosphate-dependent glutaminase was purified to homogeneity from isolated mitochondria of Ehrlich ascites-tumour cells. The enzyme had an Mr of 135,000 as judged by chromatography on Sephacryl S-300. SDS/polyacrylamide-gel electrophoresis displayed two protein bands, with Mr values of 64,000 and 56,000. Two major immunoreactive peptides of Mr values of 65,000 and 57,000 were found by immunoblot analysis using anti-(rat kidney glutaminase) antibodies. The concentration-dependences for both glutamine and phosphate were sigmoidal, with S0.5 values of 7.6 mM and 48 mM, and Hill coefficients of 1.5 and 1.6, respectively. The glutaminase pH optimum was 9. The activation energy of the enzymic reaction was 58 kJ/mol. The enzyme showed a high specificity towards glutamine. A possible explanation for the different kinetic behaviour found for purified enzyme and for isolated mitochondria [Kovacević (1974) Cancer Res. 34, 3403-3407] should be that a conformational change occurs when the enzyme is extracted from the mitochondrial inner membrane.

Nuclease sensitivity of postreplicated chromatin from Ehrlich ascites tumour cells

1981 ◽  
Vol 59 (1) ◽  
pp. 22-29
Author(s):  
Brian L. A. Miki ◽  
John J. Heikkila ◽  
Ian R. Brown
Keyword(s):  
Structural Modification ◽  
Micrococcal Nuclease ◽  
Base Pairs ◽  
Selective Labelling ◽  
Dna Repeat ◽  
Ehrlich Ascites ◽  
Nuclease Digestion ◽  
Ehrlich Ascites Tumour ◽  
Ascites Tumour Cells ◽  
Ehrlich Ascites Tumour Cells

Chromatin appears to undergo structural modification after replication and before integration into bulk chromatin. In ascites cells, postreplicated chromatin displays a transient resistance to digestion with micrococcal nuclease. This resistance may be correlated with a shorter DNA repeat length (178 base pairs) than that found in bulk chromatin (187 base pairs). Selective labelling or selective digestion of DNA sequence classes could not account for these observations. In both bulk and postreplicated chromatin, three electrophoretic types of mononucleosomes were found. Postreplicated mononucleosome types showed selective sensitivities to nuclease digestion whereas bulk mononucleosome types did not.

scholarly journals Purification and properties of a novel Ca2+-binding protein (10.5 kDa) from Ehrlich-ascites-tumour cells

1987 ◽  
Vol 247 (3) ◽  
pp. 663-667 ◽  
Author(s):  
J Kuźnicki ◽  
A Filipek
Keyword(s):  
Binding Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Tumour Cells ◽  
Ascites Tumour ◽  
Tyrosine Fluorescence ◽  
Purification And Properties ◽  
Ehrlich Ascites ◽  
Ehrlich Ascites Tumour ◽  
Ascites Tumour Cells ◽  
Ehrlich Ascites Tumour Cells

A novel Ca2+-binding protein (CaBP) was identified in Ehrlich-ascites-tumour cells and purified to homogeneity. The molecular mass of this protein is about 10.5 kDa as estimated by polyacrylamide-gel electrophoresis in the presence of SDS. CaBP has two Ca2+-binding sites that bind Ca2+ with a dissociation constant of about 3 × 10(-6)M. Ca2+ binding to CaBP decreases its electrophoretic mobility in urea/polyacrylamide gels, changes its u.v. spectrum, increases the intrinsic tyrosine fluorescence intensity and strengthens hydrophobic interaction with the phenyl-Sepharose matrix.

scholarly journals The transport and oxidation of succinate by Ehrlich ascites-tumour cells

1976 ◽  
Vol 160 (1) ◽  
pp. 121-123 ◽  
Author(s):  
T L Spencer
Keyword(s):  
Ph Dependence ◽  
Organic Anion ◽  
Tumour Cells ◽  
Cell Integrity ◽  
Carrier System ◽  
Ascites Tumour ◽  
Ehrlich Ascites ◽  
Ehrlich Ascites Tumour ◽  
Ascites Tumour Cells ◽  
Ehrlich Ascites Tumour Cells

The transport and oxidation of succinate by functionally intact Ehrlich ascites-tumour cells was investigated. On the basis of pH dependence and inhibitor sensitivity it was concluded that succinate may be transported across the cell membrane by the organic anion carrier system. Thus the ability of isolated Ehrlich cells to oxidize succinate is real, and is not necessarily a result of damage to cell integrity.

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