Interferon alters the pattern of secreted proteins from Ehrlich ascites-tumour cells

Treatment of Ehrlich ascites-tumour (EAT) cells with interferon (IFN) abolished their ability to secrete a 32 kDa protein that was secreted by growing EAT cells. These IFN-treated cells secreted two proteins (molecular masses 100 and 89 kDa as estimated by SDS/polyacrylamide-gel electrophoresis) that were not detected in two-dimensional gel electrophoresis of the culture fluid of untreated EAT cells. The sequence of 20 amino acids from the N-terminal end of the 32 kDa protein was very similar to portions of sequences of mouse proviral gag proteins.

Download Full-text

Ehrlich ascites tumour cells show tissue-specific adherence and modify their shape upon contact with embryonic fibroblasts and myotubes

Journal of Cell Science ◽

10.1242/jcs.97.3.433 ◽

1990 ◽

Vol 97 (3) ◽

pp. 433-438

Author(s):

E. Wojciak ◽

W. Korohoda

Keyword(s):

Direct Interaction ◽

Cell Spreading ◽

Ascites Tumour ◽

L1210 Cells ◽

Ehrlich Ascites ◽

Ehrlich Ascites Tumour ◽

A Cell ◽

Eat Cells ◽

Embryo Fibroblasts ◽

And Migration

Adhesiveness of Ehrlich ascites tumour (EAT) cells to glass, to mouse peritoneal membrane, living and aldehyde-fixed mouse embryo fibroblasts and chick embryo fibroblasts, myoblasts and myotubes was investigated. The ascitic EAT cells (and leukaemia L1210 cells) did not adhere to glass and peritoneum but readily adhered to embryo fibroblasts, myoblasts and myotubes. The attachment was followed by cell spreading and migration. Fixation of fibroblasts or myogenic cells with aldehydes did not prevent ascitic cells from attaching but reduced the rate of spreading. Only direct interaction of ascitic cells with embryo myoblasts or fibroblasts induced changes in tumour cell adhesiveness followed by cell spreading and locomotion. These results are discussed in relation to an observation that ascitic cells growing as a cell suspension intraperitoneally grow as a solid tumour when injected subcutaneously.

Download Full-text

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

Biochemical Journal ◽

10.1042/bj2551031 ◽

1988 ◽

Vol 255 (3) ◽

pp. 1031-1035 ◽

Cited By ~ 22

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.

Download Full-text

Characterization of mononucleosomes and associated glycoproteins from Ehrlich ascites tumour cells

Canadian Journal of Biochemistry ◽

10.1139/o80-169 ◽

1980 ◽

Vol 58 (11) ◽

pp. 1261-1269 ◽

Cited By ~ 5

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.

Download Full-text

Blood Group A and B Determinants Are Expressed on Platelet Glycoproteins IIa, IIIa, and Ib

Thrombosis and Haemostasis ◽

10.1055/s-0038-1647483 ◽

1991 ◽

Vol 65 (02) ◽

pp. 196-201 ◽

Cited By ~ 42

Author(s):

S Santoso ◽

V Kiefel ◽

C Mueller-Eckhardt

Keyword(s):

Blood Group ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Hla Class I ◽

Two Dimensional Gel Electrophoresis ◽

Blood Group A ◽

Hla Class I Molecules ◽

Sds Page ◽

Group A ◽

Molecular Masses

SummaryWe report the localization of A, B blood group determinants on intrinsic glycoproteins using anti-A, B IgG antibodies. By radioimmunoprecipitation, anti-A and anti-B precipitated three bands with apparent molecular masses in sodium dodecylsulphate- polyacrylamide gel electrophoresis (SDS-PAGE) of 159, ~ 120 and 85 kDa under non-reduced conditions and 145, ~ 126 and 97 kDa under reduced conditions. In two-dimensional gel electrophoresis (isoelectric focussing/SDS-PAGE) these three bands could be resolved into six spots and fulfilled previously defined citeria for platelet membrane glycoprotein complexes Ia/IIa, Ic’IIIa, Ib/IX and IIb/IIIa. The results were supported by data obtained by an assay employing antibody-specific immobilizatisn of platelet antigens (MAIPA). By this technique, blood group A, B determinants were shown to be immobilized by monoclonal antibodies specific for GPIa, Ic’, IX, IIb/IIIa and strongly by mab specific for GPIIa, but not by mab specific for HLA class I molecules.The more precise localization on platelet glycoproteins was achieved by immunoblotting technique by which blood group A determinants could be assigned to GPs IIa, IIIa and Ib.

Download Full-text

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

Biochemical Journal ◽

10.1042/bj2470663 ◽

1987 ◽

Vol 247 (3) ◽

pp. 663-667 ◽

Cited By ~ 58

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.

Download Full-text