Studies on the surface properties of hybrid cells. III. A membrane glycoprotein found on the surface of a wide range of malignant cells

1981 ◽  
Vol 48 (1) ◽  
pp. 147-170
Author(s):  
M.A. Atkinson ◽  
M.E. Bramwell

We report here the presence of a glycoprotein of apparent molecular mass 90000 daltons on the surface of membranes of malignant cells, which is absent or very much reduced on the surface of non-malignant cells. This glycoprotein is rich in sialic acid and appears to be sensitive to the concentration of cAMP under certain conditions. Analysis of the labelled sugars present in the glycoproteins of cells metabolically labelled with [14C]glucosamine suggests that all the enzymes necessary for the conversion of the tracer precursor into the sugars normally found to be labelled are present in both the malignant and the non-malignant cells.

1980 ◽  
Vol 46 (1) ◽  
pp. 203-220
Author(s):  
M.A. Atkinson ◽  
M.E. Bramwell

A measurable surface activity of sialyl-transferase is demonstrated by a number of different methods to exist on the plasma membranes of both malignant and non-malignant cells. The amount of enzyme present on the surface of malignant cells is found to be higher than that on the non-malignant ones in a wide range of malignant and non-malignant cell lines. It is proposed that the difference in apparent activity results in part from the presence of incomplete glycoproteins in the surface membranes of the malignant cells and in part from an increased rate of membrane synthesis in these cells.


1980 ◽  
Vol 46 (1) ◽  
pp. 187-201
Author(s):  
M.A. Atkinson ◽  
M.E. Bramwell

We report here a method for the assay of the sialyl-transferase activity in crude homogenates of a wide range of cell lines growing in tissue culture. Our results indicate that particulate preparations from both malignant and non-malignant cells show a Km of 0.25 mM towards CMP-sialic acid in the presence of an excess of glycoprotein acceptor. There appear to be increased amounts of the enzyme associated with the preparations from malignant sources which are reflected in an increase in the apparent Vmax of these. The elevated level of sialyl-transferase activity seen in the malignant cell populations is, paradoxically, associated with a decrease in the amount of bound sialic acid associated with both the whole cell homogenate preparations and the surface of these cells.


1974 ◽  
Vol 16 (1) ◽  
pp. 189-198
Author(s):  
F. WIENER ◽  
G. KLEIN ◽  
H. HARRIS

Previous studies with a variety of transplantable mouse tumours showed that in hybrids between malignant and non-malignant cells, malignancy behaved as a recessive character: the hybrid cells, so long as they retained something close to the complete parental chromosome sets, had little or no ability to grow progressively in vivo. In the experiments we now describe the heritable lesions determining the malignant phenotype were further explored by complementation analysis in which the various tumour cells were fused with each other. Forty-two clonal populations derived from twelve crosses between different kinds of tumour cells were examined. Only one cross generated hybrid cells with reduced tumorigenicity: in all other cases the hybrid cells formed were highly malignant. It thus appears that, in a wide range of different tumours, the lesions determining the malignant phenotype, although recessive, fail to complement each other.


1985 ◽  
Vol 79 (1) ◽  
pp. 83-94 ◽  
Author(s):  
H. Harris

When malignant cells, defined by their ability to grow progressively in genetically compatible hosts, are fused with diploid fibroblasts of the same species, the resulting hybrid cells, so long as they retain certain specific chromosomes donated by the diploid parent cell, are non-malignant. When these particular chromosomes are eliminated from the hybrid, the malignant phenotype reappears, and the segregant cell is again able to grow progressively in vivo. In the present experiments the histological character of the lesions produced by the inoculation of crosses between malignant and non-malignant cells was examined. It was found, in a wide range of material, and without exception, that where one or other of the parent cells in the cross was of fibroblastic lineage, malignancy was suppressed when the hybrid cells produced a collagenous extracellular matrix in vivo; and it reappeared when genetic segregants were produced that had lost the ability to produce this matrix. These results are interpreted in terms of a general model in which it is proposed that the progressive multiplication of malignant cells in vivo is a secondary consequence of a genetically stable impairment of terminal differentiation.


1990 ◽  
Vol 111 (6) ◽  
pp. 2681-2692 ◽  
Author(s):  
R J Wieser ◽  
S Schütz ◽  
G Tschank ◽  
H Thomas ◽  
H P Dienes ◽  
...  

Previous studies have shown that plasma membrane compounds are involved in the contact-dependent inhibition of growth of human diploid fibroblasts. The purification of the active plasma membrane glycoprotein is described in this report. The glycoprotein has an apparent molecular mass of 60-70 kD and, due to differential sialylation, isoelectric points between pH 5.5. and 6.2. Treatment with sialidase yielded one spot in two-dimensional gel electrophoresis with an isoelectric point of 6.3. After removal of the N-glycosidically linked oligosaccharide chains, the apparent molecular mass is reduced by approximately 22 kD. Treatment was diluted NaOH, which removes the O-glycosidically linked portion of oligosaccharides, resulted in a reduction of the apparent molecular mass by approximately 5 kD. The addition of 50 ng/ml of this glycoprotein-for which the term "contactinhibin" is proposed-in immobilized form to sparsely seeded human fibroblasts resulted in a reversible 70-80% inhibition of growth. The inhibition was not confined to human fibroblasts as other cells were also inhibited, with the exclusion of transformed cells, which are refractory to contactinhibin. The inhibitory activity was abolished by treatment with beta-galactosidase or glycopeptidase F, indicating that the glycan moiety is the biologically active part of the molecule. Confluent cultures treated with antibodies raised against contactinhibin were released from the contact-dependent inhibition of growth. In addition to enhanced saturation density, these cultures exhibited a crisscross growth pattern and the formation of foci. Immunocytochemical studies showed that contactinhibin was associated with vimentin. Furthermore, contactinhibin was found to be not expressed in a species- or organ-specific manner.


Malignancy, as measured by the ability of cells to grow progressively in vivo , is intimately linked to the presence of a structural abnormality in the polysaccharide moiety of one particular membrane glycoprotein. This abnormality is present in a wide range of different tumours; it co-segregates with malignancy in all crosses between malignant and non-malignant cells that have so far been tested; and it remains linked to malignancy in a stringent new test in which non-malignant variants are selected from tumour cell populations by the use of a lectin.


Materials ◽  
2020 ◽  
Vol 13 (10) ◽  
pp. 2295 ◽  
Author(s):  
Ryung Il Kim ◽  
Ju Ho Shin ◽  
Jong Suk Lee ◽  
Jung-Hyun Lee ◽  
Albert S. Lee ◽  
...  

A series of UV-curable hybrid composite blends containing a carboxylic acid functionalized polyimidewith varying amounts of high molecular weight (~1 K) PEG-grafted ladder-structured polysilsesquioxanes copolymerized with methacryl groups were fabricated and their structural, thermal, mechanical, and surface properties characterized. At a composite weight ratio of polyimide above 50 wt.%, a stark shift from amorphous to crystalline polyethylene glycol (PEG) phases were observed, accompanied by a drastic increase in both surface moduli and brittleness index. Moreover, fabricated composites were shown to have a wide range water contact angle, 9.8°–73.8°, attesting to the tunable surface properties of these amphiphilic hybrid polymer composites. The enhanced mechanical properties, combined with the utility of tunable surface hydrophilicity allows for the possible use of these hybrid polymer composites to be utilized as photosensitive polyimide negative photoresists for a myriad of semiconductor patterning processes.


2002 ◽  
Vol 70 (9) ◽  
pp. 4917-4924 ◽  
Author(s):  
Kleoniki Gounaris

ABSTRACT Extracellular nucleotides are signaling molecules whose receptor-mediated effects are involved in a variety of physiological responses in mammalian tissues. An overwhelming body of data indicate that inflammatory and other immune responses can be modulated by the availability and local concentrations of nucleotides via nucleotide receptor signaling, but this is only just beginning to be investigated in the context of infectious disease. Evidence is provided here that the parasitic nematode Trichinella spiralis can catalyze the conversion and thus modulate both the availability and concentration of extracellular nucleotides by means of the following secreted exoenzymes: apyrase, 5′-nucleotidase, and adenosine deaminase. These enzymes were characterized in terms of substrate specificity, kinetic behavior, pH, divalent cation preferences, and response to a series of compounds. The secreted 5′-nucleotidase was identified as a protein with an apparent molecular mass of 67 kDa after N-terminal amino acid sequencing of the purified protein. The presence of adenosine deaminase was confirmed in the secreted products by Western blotting with an antibody against a mammalian enzyme, as a protein with an apparent molecular mass of 38 kDa. These secreted proteins constitute an enzymatic cascade which catalyzes the degradation of extracellular nucleotides, with a potential physiological role in the regulation of purinergic signaling.


1999 ◽  
Vol 181 (1) ◽  
pp. 91-99 ◽  
Author(s):  
Hisayo Ono ◽  
Kazuhisa Sawada ◽  
Nonpanga Khunajakr ◽  
Tao Tao ◽  
Mihoko Yamamoto ◽  
...  

ABSTRACT 1,4,5,6-Tetrahydro-2-methyl-4-pyrimidinecarboxylic acid (ectoine) is an excellent osmoprotectant. The biosynthetic pathway of ectoine from aspartic β-semialdehyde (ASA), in Halomonas elongata, was elucidated by purification and characterization of each enzyme involved. 2,4-Diaminobutyrate (DABA) aminotransferase catalyzed reversively the first step of the pathway, conversion of ASA to DABA by transamination with l-glutamate. This enzyme required pyridoxal 5′-phosphate and potassium ions for its activity and stability. The gel filtration estimated an apparent molecular mass of 260 kDa, whereas molecular mass measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was 44 kDa. This enzyme exhibited an optimum pH of 8.6 and an optimum temperature of 25°C and had Km s of 9.1 mM forl-glutamate and 4.5 mM for dl-ASA. DABA acetyltransferase catalyzed acetylation of DABA to γ-N-acetyl-α,γ-diaminobutyric acid (ADABA) with acetyl coenzyme A and exhibited an optimum pH of 8.2 and an optimum temperature of 20°C in the presence of 0.4 M NaCl. The molecular mass was 45 kDa by gel filtration. Ectoine synthase catalyzed circularization of ADABA to ectoine and exhibited an optimum pH of 8.5 to 9.0 and an optimum temperature of 15°C in the presence of 0.5 M NaCl. This enzyme had an apparent molecular mass of 19 kDa by SDS-PAGE and a Km of 8.4 mM in the presence of 0.77 M NaCl. DABA acetyltransferase and ectoine synthase were stabilized in the presence of NaCl (>2 M) and DABA (100 mM) at temperatures below 30°C.


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