Sarcoma growth factor from mouse sarcoma virus-transformed cells. Purification by binding and elution from epidermal growth factor receptor-rich cells.

Larco, Joseph E. De; Reynolds, Roberta K.; Carlberg, Kristen; Engle, Claire G.; Todaro, George J.

doi:10.1016/s0021-9258(19)85758-7

Cited by 73 publications

(3 citation statements)

References 16 publications

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“…The results reported here indicate that exposure Of NRK cells to biologically active concentrations of TGF-/3 leads to a change in EGF/TGF-a cell surface receptors. In confirmation of previous results (39), we find that EGF/TGF-a receptors in NRK cells that are mitogenically responsive to TGF-/3 exhibit a complex ligand binding pattern (Fig. 4).…”

Section: Discussionsupporting

confidence: 92%

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Transforming growth factor-beta modulates the high-affinity receptors for epidermal growth factor and transforming growth factor-alpha.

Massagué¹

1985

The Journal of Cell Biology

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The epidermal growth factor (EGF) receptor mediates the induction of a transformed phenotype in normal rat kidney (NRK) cells by transforming growth factors (TGFs). The ability of EGF and its analogue TGF-a to induce the transformed phenotype in NRK cells is greatly potentiated by TGF-~, a polypeptide that does not interact directly with binding sites for EGF or TGF-a. Our evidence indicates that TGF-~ purified from retrovirally transformed rat embryo cells and human platelets induces a rapid (tl/2 = 0.3 h) decrease in the binding of EGF and TGF-a to high-affinity cell surface receptors in NRK cells. No change due to TGF-~ was observed in the binding of EGF or TGF-a to lower affinity sites also present in NRK cells. The effect of TGF-~ on EGF/TGF-a receptors was observed at concentrations (0.5-20 pM) similar to those at which TGF-~ is active in promoting proliferation of NRK cells in monolayer culture and semisolid medium. Affinity labeling of NRK cells and membranes by cross-linking with receptor-bound 1251-TGF-a and 12SI-EGF indicated that both factors interact with a common 170-kD receptor structure. Treatment of cells with TGF-~ decreased the intensity of affinity-labeling of this receptor structure. These data suggest that the 170 kD high-affinity receptors for EGF and TGF-a in NRK cells are a target for rapid modulation by TGF-~.

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Section: Discussionsupporting

confidence: 92%

“…5 and 6). The sharing of a common receptor species by EGF and TGF-a has been documented in other cell types as well (1,7,39,40). The decreased affinity labeling of this receptor species in TGF-/3-treated NRK cells as compared with untreated cells is consistent with it being the target for TGF-/3 regulation of TGF-a and EGF binding.…”

Section: Discussionsupporting

confidence: 56%

Transforming growth factor-beta modulates the high-affinity receptors for epidermal growth factor and transforming growth factor-alpha.

Massagué¹

1985

The Journal of Cell Biology

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“…It can be activated extracellularly by one of several ligands that share a common EGF structural motif characterized by three loops imposed by intramolecular disulfide bonds (1). Six ligands have been well characterized to date: epidermal growth factor (EGF) (2), transforming growth factor alpha (TGFR) (3), amphiregulin (AR) (4), heparin-binding EGF-like growth factor (HB-EGF) (5), betacellulin (BTC) (6), and epiregulin (7). AR, HB-EGF, and BTC possess extended N-terminal tails that, in the case of AR and HB-EGF, include heparin binding motifs.…”

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confidence: 99%

Structure−Function Studies of Ligand-Induced Epidermal Growth Factor Receptor Dimerization

et al. 1998

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We present a novel 96-well assay which we have applied to a structure-function study of epidermal growth factor receptor dimerization. The basis of the assay lies in the increased probability of EGFRs being captured as dimers by a bivalent antibody when they are immobilized in the presence of a cognate ligand. Once immobilized, the antibody acts as a tether, retaining the receptor in its dimeric state with a resultant 5-7-fold increase in binding of a radiolabeled ligand probe. When the assay was applied to members of the EGF ligand family, murine EGF, transforming growth factor alpha, and heparin-binding EGF-like growth factor were comparable with human EGF (EC50 = 2nM); betacellulin, which has a broader receptor specificity, was slightly less effective. In contrast, amphiregulin (AR1-84), which has a truncated C-tail and lacks a conserved leucine residue, was ineffective unless used at >1 microM. We further probed the involvement of the C-tail and the conserved leucine residue in receptor dimerization by comparing the activities of two genetically modified EGFs (the chimera mEGF/TGFalpha44-50 and the EGF point mutant L47A) and a C-terminally extended form of AR (AR1-90) with those of two other unrelated EGF mutants (I23T and L15A). The potency of these ligands was in the order EGF > I23T > mEGF/TGFalpha44-50 > L47A = L15A >> AR1-90 > AR1-84. Although AR was much worse than predicted from its affinity, this defect could be partially rectified by co-localization of the immobilizing antibody with heparin. Thus, it seems likely that AR cannot dimerize the EGFR unless other accessory molecules are present to stabilize its functional association with the EGFR.

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Further study of β‐tgfs released by virally transformed and non‐transformed cells

Krycève-Martinerie

Lawrence

Crochet

et al. 1985

Intl Journal of Cancer

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Chicken embryo fibroblasts sensitized by ts RSV respond to TGFs present in the media of non-transformed FR3T3 and NRK-4 rat cells and of the same cells transformed by KiMSV or RSV. They also respond to TGFs present in the media of BHK hamster cells transformed by MoMSV, PyV or RSV. Two other indicator rat cell lines, untransformed NRK-4 and FR3T3, sensitized by ts KiMSV, respond to the same TGF-containing media, and this response is increased by exogenous EGF. Normal FR3T3 cells failed to respond to any of the media. The most sensitive target cells were the ts KiMSV-FR3T3 cells at the restrictive temperature (39.5 degrees C). All the media tested on NRK-49F target cells required EGF for their TGF activity which was essentially dependent on prior activation by acidification. These data show that the above media from non-transformed or transformed cells contain beta-TGFs, with no detectable accompanying alpha-TGF activity. The release of and the response to these TGFs are not interdependent. A function of ts mutant src and k-ras viral oncogenes, still expressed at the restrictive temperature, can sensitize non-responsive cells, without there being any specificity towards the TGF producer cells.

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Sarcoma growth factor from mouse sarcoma virus-transformed cells. Purification by binding and elution from epidermal growth factor receptor-rich cells.

Cited by 73 publications

References 16 publications

Transforming growth factor-beta modulates the high-affinity receptors for epidermal growth factor and transforming growth factor-alpha.

Transforming growth factor-beta modulates the high-affinity receptors for epidermal growth factor and transforming growth factor-alpha.

Structure−Function Studies of Ligand-Induced Epidermal Growth Factor Receptor Dimerization

Further study of β‐tgfs released by virally transformed and non‐transformed cells

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