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. 1985 Nov 1;101(5):1690–1694. doi: 10.1083/jcb.101.5.1690

Wheat germ agglutinin blocks the biological effects of nerve growth factor

PMCID: PMC2113952  PMID: 3840487

Abstract

The binding of nerve growth factor (NGF) to specific cell surface receptors initiates a variety of effects that lead to the morphological and biochemical differentiation of clonal pheochromocytoma, PC12, cells. The lectin wheat germ agglutinin (WGA) alters the characteristics of NGF-receptor interaction. We have found that treatment of PC12 cells with WGA dramatically and reversibly inhibits the ability of NGF to elicit three distinct biological effects characteristic of NGF action. Two of these events, the rapid ruffling of cell-surface membranes and the stimulation of the phosphorylation of a 250-kD cytoskeletal protein in situ, occur rapidly and are an immediate consequence of receptor occupancy. Both of these effects are blocked by pretreatment of the cells with WGA. WGA was also found to inhibit the NGF-stimulated regeneration of neurites that occurs over 1- 2 d. Both the WGA inhibition of neurite outgrowth and the phosphorylation of the 250-kD cytoskeletal protein were reversed upon addition of the specific sugar N-acetylglucosamine. These data demonstrate that the WGA-induced changes in the NGF-receptor interaction reflect important alterations in the ability of the receptor to transmit biological signals, resulting in the abrogation of the biological effects of NGF on these cells.

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Selected References

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  1. Bernd P., Greene L. A. Association of 125I-nerve growth factor with PC12 pheochromocytoma cells. Evidence for internalization via high-affinity receptors only and for long-term regulation by nerve growth factor of both high- and low-affinity receptors. J Biol Chem. 1984 Dec 25;259(24):15509–15516. [PubMed] [Google Scholar]
  2. Block T., Bothwell M. The nerve growth factor receptor on PC12 cells: interconversion between two forms with different binding properties. J Neurochem. 1983 Jun;40(6):1654–1663. doi: 10.1111/j.1471-4159.1983.tb08139.x. [DOI] [PubMed] [Google Scholar]
  3. Buxser S. E., Kelleher D. J., Watson L., Puma P., Johnson G. L. Change in state of nerve growth factor receptor. Modulation of receptor affinity by wheat germ agglutinin. J Biol Chem. 1983 Mar 25;258(6):3741–3749. [PubMed] [Google Scholar]
  4. Buxser S. E., Watson L., Johnson G. L. A comparison of binding properties and structure of NGF receptor on PC12 pheochromocytoma and A875 melanoma cells. J Cell Biochem. 1983;22(4):219–233. doi: 10.1002/jcb.240220404. [DOI] [PubMed] [Google Scholar]
  5. Buxser S., Puma P., Johnson G. L. Properties of the nerve growth factor receptor. Relationship between receptor structure and affinity. J Biol Chem. 1985 Feb 10;260(3):1917–1926. [PubMed] [Google Scholar]
  6. Connolly J. L., Greene L. A., Viscarello R. R., Riley W. D. Rapid, sequential changes in surface morphology of PC12 pheochromocytoma cells in response to nerve growth factor. J Cell Biol. 1979 Sep;82(3):820–827. doi: 10.1083/jcb.82.3.820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Green L. A. A quantitative bioassay for nerve growth factor (NGF) activity employing a clonal pheochromocytoma cell line. Brain Res. 1977 Sep 16;133(2):350–353. doi: 10.1016/0006-8993(77)90770-3. [DOI] [PubMed] [Google Scholar]
  8. Greene L. A., Shooter E. M. The nerve growth factor: biochemistry, synthesis, and mechanism of action. Annu Rev Neurosci. 1980;3:353–402. doi: 10.1146/annurev.ne.03.030180.002033. [DOI] [PubMed] [Google Scholar]
  9. Greene L. A., Tischler A. S. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2424–2428. doi: 10.1073/pnas.73.7.2424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grob P. M., Berlot C. H., Bothwell M. A. Affinity labeling and partial purification of nerve growth factor receptors from rat pheochromocytoma and human melanoma cells. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6819–6823. doi: 10.1073/pnas.80.22.6819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grob P. M., Bothwell M. A. Modification of nerve growth factor receptor properties by wheat germ agglutinin. J Biol Chem. 1983 Dec 10;258(23):14136–14143. [PubMed] [Google Scholar]
  12. Hosang M., Shooter E. M. Molecular characteristics of nerve growth factor receptors on PC12 cells. J Biol Chem. 1985 Jan 10;260(1):655–662. [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Landreth G. E., Rieser G. D. Nerve growth factor- and epidermal growth factor-stimulated phosphorylation of a PC12 cytoskeletally associated protein in situ. J Cell Biol. 1985 Mar;100(3):677–683. doi: 10.1083/jcb.100.3.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Landreth G. E., Shooter E. M. Nerve growth factor receptors on PC12 cells: ligand-induced conversion from low- to high-affinity states. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4751–4755. doi: 10.1073/pnas.77.8.4751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Massague J., Buxser S., Johnson G. L., Czech M. P. Affinity labeling of a nerve growth factor receptor component on rat pheochromocytoma (PC12) cells. Biochim Biophys Acta. 1982 Dec 8;693(1):205–212. doi: 10.1016/0005-2736(82)90488-6. [DOI] [PubMed] [Google Scholar]
  17. Puma P., Buxser S. E., Watson L., Kelleher D. J., Johnson G. L. Purification of the receptor for nerve growth factor from A875 melanoma cells by affinity chromatography. J Biol Chem. 1983 Mar 10;258(5):3370–3375. [PubMed] [Google Scholar]
  18. Ross A. H., Grob P., Bothwell M., Elder D. E., Ernst C. S., Marano N., Ghrist B. F., Slemp C. C., Herlyn M., Atkinson B. Characterization of nerve growth factor receptor in neural crest tumors using monoclonal antibodies. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6681–6685. doi: 10.1073/pnas.81.21.6681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schechter A. L., Bothwell M. A. Nerve growth factor receptors on PC12 cells: evidence for two receptor classes with differing cytoskeletal association. Cell. 1981 Jun;24(3):867–874. doi: 10.1016/0092-8674(81)90112-4. [DOI] [PubMed] [Google Scholar]
  20. Smith A. P., Varon S., Shooter E. M. Multiple forms of the nerve growth factor protein and its subunits. Biochemistry. 1968 Sep;7(9):3259–3268. doi: 10.1021/bi00849a032. [DOI] [PubMed] [Google Scholar]
  21. Thoenen H., Barde Y. A. Physiology of nerve growth factor. Physiol Rev. 1980 Oct;60(4):1284–1335. doi: 10.1152/physrev.1980.60.4.1284. [DOI] [PubMed] [Google Scholar]
  22. Vale R. D., Shooter E. M. Alteration of binding properties and cytoskeletal attachment of nerve growth factor receptors in PC12 cells by wheat germ agglutinin. J Cell Biol. 1982 Sep;94(3):710–717. doi: 10.1083/jcb.94.3.710. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Vale R. D., Shooter E. M. Epidermal growth factor receptors on PC12 cells: alteration of binding properties by lectins. J Cell Biochem. 1983;22(2):99–109. doi: 10.1002/jcb.240220204. [DOI] [PubMed] [Google Scholar]

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