Mitogenic proteins of the 3T3 plasma membrane

Lieberman, M. A.

doi:10.1002/jcp.1041140112

Cited by 16 publications

(10 citation statements)

References 25 publications

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“…A 150,000-Da trypsin-releasable mitogenic activity associated with 3T3 plasma membranes has been reported by Lieberman (18). The active component described here is much smaller than this factor and is different in its potency, acid resistance, and DEAE-cellulose elution characteristics.…”

supporting

confidence: 48%

Identification of a 34,000-dalton mitogenic protein associated with plasma membranes from human A431 epidermoid carcinoma cells.

Bishayee¹,

Matesic²,

Das³

1984

Proc. Natl. Acad. Sci. U.S.A.

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We present evidence for a discrete 34,000-Da polypeptide with mitogenic activity, associated with plasma membranes from human A431 carcinoma cells. Plasma The role of hormones and growth factors in animal cell proliferation is well established, and many aspects of the interaction of these factors with responsive cells have been worked out (1-5). However, growth regulation by cell-cell contact is poorly understood. It is believed that surface contact between cells is partly responsible for density-dependent inhibition of growth (6, 7). Again, under certain circumstances surface contact can be stimulatory (for example, contact with axonal membranes is known to stimulate Schwann cell division) (8). Thus, cell-cell interaction may generate either mitogenic or antimitogenic signals. In no case has the molecular basis for these growth regulatory effects been identified.In an earlier communication, we reported the properties of a mitogenic activity associated with mouse hepatic membranes (9, 10). The activity was found to be protease sensitive and heat labile. We now find that the plasma membrane from A431 cells, a human epidermoid carcinoma line, is an unusually potent mitogen for target fibroblastic cells. Approximately 30% of the membrane-associated activity can be solubilized with high salt concentration in the absence of detergents, and this apparently represents peripherally attached membrane components. Incubation of this fraction with target fibroblast cells results in selective binding of a single protein of 34,000 Da.These fortunate characteristics-the high mitogenic potency of A431 membranes, the salt extractability of a fraction of this activity, and the selective binding of a particular protein from this fraction by target cells-made possible the studies described here. Evidence so far obtained indicates that the 34,000-Da polypeptide represents a peripherally attached cell-surface mitogen. MATERIALS AND METHODSCell Culture and Plasma Membranes. Monolayer cultures of Swiss mouse NR-6 cells (11,12) and human A431 cells were grown in Dulbecco's modified Eagle's (DME) medium containing 7% fetal bovine serum and gentamycin at 10 .tg/ml. Membranes were prepared from A431 cells using the procedure of Thom et al. (13).Extract of A431 Membranes. Washed A431 plasma membrane (10 mg of protein) was suspended in 8 ml of 0.7 M NaCl/10 mM Hepes, pH 7.4, and stirred at 40C for 2 hr. The suspension was centrifuged at 100,000 x g for 60 min, and the supernate was concentrated by Amicon ultrafiltration using PM-10 filters. During ultrafiltration, the NaCl concentration was adjusted to 0.15 M.Acetic Acid Treatment of Extract. To extract of A431 membranes treated with a high concentration of salt (1 mg of protein per ml of 0.15 M NaCl/10 mM Hepes, pH 7.4), acetic acid was added to a final concentration of 0.34 M. After incubation at 4°C for 30 min, the mixture was neutralized with saturated Tris solution, and clarified by centrifugation at 100,000 X g for 60 min.Incorporation of [3H]Thymidine into Acid-Insoluble Mat...

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supporting

confidence: 48%

Identification of a 34,000-dalton mitogenic protein associated with plasma membranes from human A431 epidermoid carcinoma cells.

Bishayee¹,

Matesic²,

Das³

1984

Proc. Natl. Acad. Sci. U.S.A.

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“…In this report, we describe the observation that heparin can also potentiate the action of plasma membraneassociated growth stimulatory activity (PMGA), which is found in a high-salt extract of cellular plasma membranes (Lieberman, 1983(Lieberman, ,1984. By using a pyrophosphate extract of mouse liver plasma membranes, we have found that the action of heparin can be mimicked by several sulfated polysaccharides.…”

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confidence: 89%

Heparin potentiates the action of plasma membrane‐associated growth stimulatory activity

Nagasaki

Lieberman

1987

Journal Cellular Physiology

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Plasma membranes prepared from mouse liver have been previously shown to contain growth stimulatory activity as determined with cultured mouse fibroblasts. This growth stimulatory activity, termed plasma membrane-associated growth stimulatory activity (PMGA), is highly mitogenic in the presence of platelet-poor plasma. We now demonstrate that the growth stimulatory action of PMGA is dramatically enhanced by the addition of heparin. The half-maximal effect of heparin was observed at 1-3 micrograms/ml. The synergistic effect was seen in two distinct assays; the stimulation of DNA synthesis in quiescent cells, and an increase of cell number over a 3-day culture period. Heparin, by itself, does not have any measurable influence on the growth of fibroblasts. The action of heparin is not unique to this glycosaminoglycan, as several other highly sulfated polysaccharides, including dextran sulfate, pentosan polysulfate, and fucoidan, also exhibited the highly synergistic effect. Among other glycosaminoglycans examined, chondroitin sulfate B and heparan sulfate had a small, but significant, effect on enhancing the growth stimulatory action of PMGA. Chondroitin sulfate A, chondroitin sulfate C, hyaluronic acid dextran, and poly-L-glutamic acid, however, had no detectable effect. Further experiments suggested that the effect of heparin is twofold, namely, both a potentiation of growth stimulatory activity and a protection of PMGA activity. The data presented here suggest that the association of various cell surface components, such as PMGA and specific proteoglycans, can modulate the growth potential of a cell.

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“…The next group of growth inhibitors encompasses peptide factors isolated from plasma membrane fractions of resting cultured cells (Whittenberger & Glaser 1977, Datta & Natraj 1980, Lieberman 1983. Similar fractions from growing cells lack the inhibitory activity.…”

Section: Cell-associated Inhibitorsmentioning

confidence: 99%

“…A human cdc2.5 homologue essential for mitosis has also been identified (Millar et al 1991, Strausfeld et al 1991. Conversely, the products of two genes, weel and mikl (both kinases) are negative regulators of ~3 4 '~" " , phosphorylating it on tyrosine residues and thus preventing the onset of mitosis (Lundgren et al 1991). The negative effects of the weel gene can be overridden by the product of another gene, niml kinase (Russell & Nurse 1987, Wu & Russell 1993, Parker et al 1993, whereas the products of the pypl and pyp2 genes (both tyrosine phosphatases) contribute to the inhibition of mitosis by enhancing the activity of weel kinase (Millar et al 1992).…”

Section: T H E G/m Transitionmentioning

confidence: 99%

Negative control of cell proliferation in eukaryotes

Epifanova

Brooks

1994

Cell Proliferation

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The nucleoside diphosphate kinase (NDPK/nm23) iso-forms H1 and H2 were localized in hematopoietic tissues. Flow cytometric analysis and enzymatic assays were used to quantify the intracellular and extracellu-lar concentrations of NDPK. Bone marrow CD34 progenitors contained the highest intracellular levels of both nm23-H1 and nm23-H2. Lower levels were measured in more mature bone marrow cells, whereas peripheral blood leukocytes had the lowest expression of nm23. These data suggest a function of NDPK in early hematopoiesis and a down-regulation of NDPK upon differentiation. In addition, an up-regulation of nm23 expression was observed in lymphocytes after induction of proliferation with phytohemagglutinin. Multiparam-eter flow cytometry demonstrated that this up-regulation occurred during the G 0 /G 1-transition. Flow cyto-metric analysis also revealed a weak surface expression of nm23 on a number of hematopoietic cell lines, which was not detected on normal hematopoietic cells. Our data also demonstrated the presence of NDPK in human plasma, probably due to a limited in vivo lysis of red blood cells. Currently, the family of nucleoside diphosphate kinases (NDPK/nm23) 1 consists of four members encoded by the genes nm23-H1, nm23-H2, DR-nm23, and nm23-H4 (1-4). These enzymes share the ability to transfer the terminal phosphate of nucleoside triphosphates to nucleoside diphos-phates. Beside their function to maintain the cellular GTP level (5), several other functions and properties have been assigned to the NDPK family. These include the identity of nm23-H2 to PuF, a transcription factor of the proto-oncogene c-myc (6), the ability of nm23-H1 to suppress the metastatic potential of several tumors and cell lines (1, 7, 8), the association of NDPK with transcription regulating factors such as the retinoic acid receptor-related orphan receptor and the retinoic Z receptor , and with the heat shock protein hsc70 (9, 10). Recently, it was also reported that NDPK has protein kinase activity (11, 12). The importance of nm23 expression in hematopoiesis has been demonstrated by Okabe-Kado et al. (13), who identified NDPK B as a differentiation inhibiting factor (I-factor) in the conditioned medium of the differentiation-resistant mouse M1 cell line and demonstrated that both nm23-H1 and nm23-H2 can inhibit the in vitro induced differentiation of several hema-topoietic cell lines. This inhibition was independent of the phosphotransferase activity as demonstrated with nm23 mutants lacking the enzymatic activity (13-15). Moreover, reported data indicated a correlation between proliferation and nm23 expression in hematopoietic cells. In lym-phoma and monocytic leukemia, overexpression of nm23-H1 was correlated with a higher tumor aggressiveness and resistance to chemotherapy, respectively (16, 17). Up-regulation of nm23-H1 and-H2 was observed in normal lymphocytes induced to proliferate with phytohemagglutinin (PHA) (18, 19). In vitro, DR-nm23 mRNA is up-regulated in CD34 hemato-poietic progenitor cells during early stages of...

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Mitogenic proteins of the 3T3 plasma membrane

Cited by 16 publications

References 25 publications

Identification of a 34,000-dalton mitogenic protein associated with plasma membranes from human A431 epidermoid carcinoma cells.

Identification of a 34,000-dalton mitogenic protein associated with plasma membranes from human A431 epidermoid carcinoma cells.

Heparin potentiates the action of plasma membrane‐associated growth stimulatory activity

Negative control of cell proliferation in eukaryotes

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