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. 1989 Aug;9(8):3538–3542. doi: 10.1128/mcb.9.8.3538

Immunochemical localization of the epitope for a monoclonal antibody that neutralizes human platelet-derived growth factor mitogenic activity.

W J LaRochelle 1, K C Robbins 1, S A Aaronson 1
PMCID: PMC362402  PMID: 2477688

Abstract

A monoclonal antibody (mAb), sis 1, generated against human c-sis-encoded platelet-derived growth factor (PDGF) BB, was shown by enzyme-linked immunosorbent assay and Western blot (immunoblot) analysis to recognize human PDGF BB and human platelet PDGF AB but not the human PDGF AA. This monoclonal antibody potently inhibited PDGF receptor-binding and mitogenic activities of both human PDGF BB and PDGF AB but had no effect on PDGF AA. Finally, we demonstrated that an immunoaffinity-purified anti-c-sis peptide antibody (anti-V4) which also blocked binding of PDGF BB to its cognate receptor and competed with mAb sis 1 for binding to PDGF BB. All of these results suggest that mAb sis 1 recognizes an epitope of the c-sis gene product, PDGF BB, that spatially overlaps the V4 surface domain of PDGF BB, immunochemically localizing a region of PDGF BB critical for PDGF receptor binding and activation.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beckmann M. P., Betsholtz C., Heldin C. H., Westermark B., Di Marco E., Di Fiore P. P., Robbins K. C., Aaronson S. A. Comparison of biological properties and transforming potential of human PDGF-A and PDGF-B chains. Science. 1988 Sep 9;241(4871):1346–1349. doi: 10.1126/science.2842868. [DOI] [PubMed] [Google Scholar]
  2. Bringman T. S., Lindquist P. B., Derynck R. Different transforming growth factor-alpha species are derived from a glycosylated and palmitoylated transmembrane precursor. Cell. 1987 Feb 13;48(3):429–440. doi: 10.1016/0092-8674(87)90194-2. [DOI] [PubMed] [Google Scholar]
  3. Devare S. G., Reddy E. P., Law J. D., Robbins K. C., Aaronson S. A. Nucleotide sequence of the simian sarcoma virus genome: demonstration that its acquired cellular sequences encode the transforming gene product p28sis. Proc Natl Acad Sci U S A. 1983 Feb;80(3):731–735. doi: 10.1073/pnas.80.3.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Doolittle R. F., Hunkapiller M. W., Hood L. E., Devare S. G., Robbins K. C., Aaronson S. A., Antoniades H. N. Simian sarcoma virus onc gene, v-sis, is derived from the gene (or genes) encoding a platelet-derived growth factor. Science. 1983 Jul 15;221(4607):275–277. doi: 10.1126/science.6304883. [DOI] [PubMed] [Google Scholar]
  5. Ey P. L., Prowse S. J., Jenkin C. R. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-sepharose. Immunochemistry. 1978 Jul;15(7):429–436. doi: 10.1016/0161-5890(78)90070-6. [DOI] [PubMed] [Google Scholar]
  6. Giese N. A., Robbins K. C., Aaronson S. A. The role of individual cysteine residues in the structure and function of the v-sis gene product. Science. 1987 Jun 5;236(4806):1315–1318. doi: 10.1126/science.3035718. [DOI] [PubMed] [Google Scholar]
  7. Hannink M., Sauer M. K., Donoghue D. J. Deletions in the C-terminal coding region of the v-sis gene: dimerization is required for transformation. Mol Cell Biol. 1986 Apr;6(4):1304–1314. doi: 10.1128/mcb.6.4.1304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Igarashi H., Rao C. D., Siroff M., Leal F., Robbins K. C., Aaronson S. A. Detection of PDGF-2 homodimers in human tumor cells. Oncogene. 1987 Mar;1(1):79–85. [PubMed] [Google Scholar]
  9. Johnsson A., Heldin C. H., Westermark B., Wasteson A. Platelet-derived growth factor: identification of constituent polypeptide chains. Biochem Biophys Res Commun. 1982 Jan 15;104(1):66–74. doi: 10.1016/0006-291x(82)91941-6. [DOI] [PubMed] [Google Scholar]
  10. King C. R., Giese N. A., Robbins K. C., Aaronson S. A. In vitro mutagenesis of the v-sis transforming gene defines functional domains of its growth factor-related product. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5295–5299. doi: 10.1073/pnas.82.16.5295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kuo L. M., Robb R. J. Structure-function relationships for the IL 2-receptor system. I. Localization of a receptor binding site on IL 2. J Immunol. 1986 Sep 1;137(5):1538–1543. [PubMed] [Google Scholar]
  12. LaRochelle W. J., Wray B. E., Sealock R., Froehner S. C. Immunochemical demonstration that amino acids 360-377 of the acetylcholine receptor gamma-subunit are cytoplasmic. J Cell Biol. 1985 Mar;100(3):684–691. doi: 10.1083/jcb.100.3.684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Leal F., Williams L. T., Robbins K. C., Aaronson S. A. Evidence that the v-sis gene product transforms by interaction with the receptor for platelet-derived growth factor. Science. 1985 Oct 18;230(4723):327–330. doi: 10.1126/science.2996133. [DOI] [PubMed] [Google Scholar]
  14. Rao C. D., Igarashi H., Chiu I. M., Robbins K. C., Aaronson S. A. Structure and sequence of the human c-sis/platelet-derived growth factor 2 (SIS/PDGF2) transcriptional unit. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2392–2396. doi: 10.1073/pnas.83.8.2392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Robbins K. C., Devare S. G., Reddy E. P., Aaronson S. A. In vivo identification of the transforming gene product of simian sarcoma virus. Science. 1982 Dec 10;218(4577):1131–1133. doi: 10.1126/science.6293053. [DOI] [PubMed] [Google Scholar]
  16. Ross R., Raines E. W., Bowen-Pope D. F. The biology of platelet-derived growth factor. Cell. 1986 Jul 18;46(2):155–169. doi: 10.1016/0092-8674(86)90733-6. [DOI] [PubMed] [Google Scholar]
  17. Sauer M. K., Donoghue D. J. Identification of nonessential disulfide bonds and altered conformations in the v-sis protein, a homolog of the B chain of platelet-derived growth factor. Mol Cell Biol. 1988 Mar;8(3):1011–1018. doi: 10.1128/mcb.8.3.1011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Waterfield M. D., Scrace G. T., Whittle N., Stroobant P., Johnsson A., Wasteson A., Westermark B., Heldin C. H., Huang J. S., Deuel T. F. Platelet-derived growth factor is structurally related to the putative transforming protein p28sis of simian sarcoma virus. Nature. 1983 Jul 7;304(5921):35–39. doi: 10.1038/304035a0. [DOI] [PubMed] [Google Scholar]

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