Protamine enhances epidermal growth factor (EGF)-stimulated mitogenesis by increasing cell surface EGF receptor number. Implications for existence of cryptic EGF receptors

Vinata B Lokeshwar, S. S. Huang, J. S. Huang

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Abstract

Treatment of Swiss mouse 3T3 cells and human epidermoid carcinoma A431 cells with protamine at 37°C increased the 125I-epidermal growth factor (EGF) binding activity at 4°C. The effect of protamine on the increase of 125-EGF binding activity appeared to be time, temperature, and dose dependent. This up-modulation of 125I-EGF binding by protamine correlated with protamine enhancement of EGF-stimulated mitogenesis, with respect to the magnitude of the effect and the dose response curves. Scatchard plot analyses indicated that protamine induced an increase in numbers of both high and low affinity EGF receptors without affecting their affinities. Protamine also increased functionally active EGF receptors in plasma membranes and solubilized membranes. This was evidenced by Scatchard plot analyses and by a protamine-induced increase of 125I-EGF-EGF receptor complex and an increase in EGF-stimulated phosphorylation of the EGF receptor. Combined with column chromatography of the solubilized EGF receptor on protamine-agarose gel, these results suggest that protamine may increase the EGF receptor number by directly activating cryptic EGF receptors in the plasma membrane.

Original languageEnglish (US)
Pages (from-to)19318-19326
Number of pages9
JournalJournal of Biological Chemistry
Volume264
Issue number32
StatePublished - Dec 1 1989
Externally publishedYes

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Protamines
Epidermal Growth Factor Receptor
Epidermal Growth Factor
Cell membranes
Cell Membrane
Swiss 3T3 Cells
ErbB Receptors
Column chromatography
Phosphorylation
Sepharose
Chromatography
Squamous Cell Carcinoma
Gels
Modulation
Membranes
Temperature

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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abstract = "Treatment of Swiss mouse 3T3 cells and human epidermoid carcinoma A431 cells with protamine at 37°C increased the 125I-epidermal growth factor (EGF) binding activity at 4°C. The effect of protamine on the increase of 125-EGF binding activity appeared to be time, temperature, and dose dependent. This up-modulation of 125I-EGF binding by protamine correlated with protamine enhancement of EGF-stimulated mitogenesis, with respect to the magnitude of the effect and the dose response curves. Scatchard plot analyses indicated that protamine induced an increase in numbers of both high and low affinity EGF receptors without affecting their affinities. Protamine also increased functionally active EGF receptors in plasma membranes and solubilized membranes. This was evidenced by Scatchard plot analyses and by a protamine-induced increase of 125I-EGF-EGF receptor complex and an increase in EGF-stimulated phosphorylation of the EGF receptor. Combined with column chromatography of the solubilized EGF receptor on protamine-agarose gel, these results suggest that protamine may increase the EGF receptor number by directly activating cryptic EGF receptors in the plasma membrane.",
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AU - Lokeshwar, Vinata B

AU - Huang, S. S.

AU - Huang, J. S.

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N2 - Treatment of Swiss mouse 3T3 cells and human epidermoid carcinoma A431 cells with protamine at 37°C increased the 125I-epidermal growth factor (EGF) binding activity at 4°C. The effect of protamine on the increase of 125-EGF binding activity appeared to be time, temperature, and dose dependent. This up-modulation of 125I-EGF binding by protamine correlated with protamine enhancement of EGF-stimulated mitogenesis, with respect to the magnitude of the effect and the dose response curves. Scatchard plot analyses indicated that protamine induced an increase in numbers of both high and low affinity EGF receptors without affecting their affinities. Protamine also increased functionally active EGF receptors in plasma membranes and solubilized membranes. This was evidenced by Scatchard plot analyses and by a protamine-induced increase of 125I-EGF-EGF receptor complex and an increase in EGF-stimulated phosphorylation of the EGF receptor. Combined with column chromatography of the solubilized EGF receptor on protamine-agarose gel, these results suggest that protamine may increase the EGF receptor number by directly activating cryptic EGF receptors in the plasma membrane.

AB - Treatment of Swiss mouse 3T3 cells and human epidermoid carcinoma A431 cells with protamine at 37°C increased the 125I-epidermal growth factor (EGF) binding activity at 4°C. The effect of protamine on the increase of 125-EGF binding activity appeared to be time, temperature, and dose dependent. This up-modulation of 125I-EGF binding by protamine correlated with protamine enhancement of EGF-stimulated mitogenesis, with respect to the magnitude of the effect and the dose response curves. Scatchard plot analyses indicated that protamine induced an increase in numbers of both high and low affinity EGF receptors without affecting their affinities. Protamine also increased functionally active EGF receptors in plasma membranes and solubilized membranes. This was evidenced by Scatchard plot analyses and by a protamine-induced increase of 125I-EGF-EGF receptor complex and an increase in EGF-stimulated phosphorylation of the EGF receptor. Combined with column chromatography of the solubilized EGF receptor on protamine-agarose gel, these results suggest that protamine may increase the EGF receptor number by directly activating cryptic EGF receptors in the plasma membrane.

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