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. 1988 May;7(5):1387–1393. doi: 10.1002/j.1460-2075.1988.tb02955.x

Binding of different dimeric forms of PDGF to human fibroblasts: evidence for two separate receptor types.

C H Heldin 1, G Bäckström 1, A Ostman 1, A Hammacher 1, L Rönnstrand 1, K Rubin 1, M Nistér 1, B Westermark 1
PMCID: PMC458388  PMID: 2842148

Abstract

The binding of the three dimeric forms of platelet-derived growth factor (PDGF), PDGF-AA, PDGF-AB and PDGF-BB, to human fibroblasts was studied. Cross-competition experiments revealed the existence of two different PDGF receptor classes: the type A PDGF receptor bound all three dimeric forms of PDGF, whereas the type B PDGF receptor bound PDGF-BB with high affinity and PDGF-AB with lower affinity, but not PDGF-AA. The sizes of the two receptors were estimated with affinity labeling techniques; the A type receptor appeared as a major component of 125 kd and a minor of 160 kd, and the B type receptor as two components of 160 and 175 kd. A previously established PDGF receptor monoclonal antibody, PDGFR-B2, was shown to react with the B type receptor only. The different abilities of the three dimeric forms of PDGF to stimulate incorporation of [3H]TdR into human fibroblasts indicated that the major mitogenic effect of PDGF is mediated via the B type receptor.

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Selected References

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  1. Betsholtz C., Johnsson A., Heldin C. H., Westermark B., Lind P., Urdea M. S., Eddy R., Shows T. B., Philpott K., Mellor A. L. cDNA sequence and chromosomal localization of human platelet-derived growth factor A-chain and its expression in tumour cell lines. Nature. 1986 Apr 24;320(6064):695–699. doi: 10.1038/320695a0. [DOI] [PubMed] [Google Scholar]
  2. Betsholtz C., Westermark B. Growth factor-induced proliferation of human fibroblasts in serum-free culture depends on cell density and extracellular calcium concentration. J Cell Physiol. 1984 Feb;118(2):203–210. doi: 10.1002/jcp.1041180213. [DOI] [PubMed] [Google Scholar]
  3. Bishayee S., Ross A. H., Womer R., Scher C. D. Purified human platelet-derived growth factor receptor has ligand-stimulated tyrosine kinase activity. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6756–6760. doi: 10.1073/pnas.83.18.6756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blobel G., Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. doi: 10.1083/jcb.67.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bolton A. E., Hunter W. M. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J. 1973 Jul;133(3):529–539. doi: 10.1042/bj1330529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Claesson-Welsh L., Rönnstrand L., Heldin C. H. Biosynthesis and intracellular transport of the receptor for platelet-derived growth factor. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8796–8800. doi: 10.1073/pnas.84.24.8796. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Daniel T. O., Tremble P. M., Frackelton A. R., Jr, Williams L. T. Purification of the platelet-derived growth factor receptor by using an anti-phosphotyrosine antibody. Proc Natl Acad Sci U S A. 1985 May;82(9):2684–2687. doi: 10.1073/pnas.82.9.2684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davis R. J., Czech M. P. Platelet-derived growth factor mimics phorbol diester action on epidermal growth factor receptor phosphorylation at threonine-654. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4080–4084. doi: 10.1073/pnas.82.12.4080. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Glenn K., Bowen-Pope D. F., Ross R. Platelet-derived growth factor. III. Identification of a platelet-derived growth factor receptor by affinity labeling. J Biol Chem. 1982 May 10;257(9):5172–5176. [PubMed] [Google Scholar]
  11. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  12. Hart C. E., Seifert R. A., Ross R., Bowen-Pope D. F. Synthesis, phosphorylation, and degradation of multiple forms of the platelet-derived growth factor receptor studied using a monoclonal antibody. J Biol Chem. 1987 Aug 5;262(22):10780–10785. [PubMed] [Google Scholar]
  13. Heldin C. H., Ek B., Rönnstrand L. Characterization of the receptor for platelet-derived growth factor on human fibroblasts. Demonstration of an intimate relationship with a 185,000-Dalton substrate for the platelet-derived growth factor-stimulated kinase. J Biol Chem. 1983 Aug 25;258(16):10054–10061. [PubMed] [Google Scholar]
  14. Heldin C. H., Johnsson A., Ek B., Wennergren S., Rönnstrand L., Hammacher A., Faulders B., Wasteson A., Westermark B. Purification of human platelet-derived growth factor. Methods Enzymol. 1987;147:3–13. doi: 10.1016/0076-6879(87)47094-8. [DOI] [PubMed] [Google Scholar]
  15. Heldin C. H., Johnsson A., Wennergren S., Wernstedt C., Betsholtz C., Westermark B. A human osteosarcoma cell line secretes a growth factor structurally related to a homodimer of PDGF A-chains. Nature. 1986 Feb 6;319(6053):511–514. doi: 10.1038/319511a0. [DOI] [PubMed] [Google Scholar]
  16. Heldin C. H., Wasteson A., Westermark B. Interaction of platelet-derived growth factor with its fibroblast receptor. Demonstration of ligand degradation and receptor modulation. J Biol Chem. 1982 Apr 25;257(8):4216–4221. [PubMed] [Google Scholar]
  17. Heldin C. H., Wasteson A., Westermark B. Platelet-derived growth factor. Mol Cell Endocrinol. 1985 Mar;39(3):169–187. doi: 10.1016/0303-7207(85)90061-9. [DOI] [PubMed] [Google Scholar]
  18. Johnsson A., Heldin C. H., Wasteson A., Westermark B., Deuel T. F., Huang J. S., Seeburg P. H., Gray A., Ullrich A., Scrace G. The c-sis gene encodes a precursor of the B chain of platelet-derived growth factor. EMBO J. 1984 May;3(5):921–928. doi: 10.1002/j.1460-2075.1984.tb01908.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Josephs S. F., Guo C., Ratner L., Wong-Staal F. Human-proto-oncogene nucleotide sequences corresponding to the transforming region of simian sarcoma virus. Science. 1984 Feb 3;223(4635):487–491. doi: 10.1126/science.6318322. [DOI] [PubMed] [Google Scholar]
  20. Keating M. T., Williams L. T. Processing of the platelet-derived growth factor receptor. Biosynthetic and degradation studies using anti-receptor antibodies. J Biol Chem. 1987 Jun 5;262(16):7932–7937. [PubMed] [Google Scholar]
  21. Markwell M. A. A new solid-state reagent to iodinate proteins. I. Conditions for the efficient labeling of antiserum. Anal Biochem. 1982 Sep 15;125(2):427–432. doi: 10.1016/0003-2697(82)90025-2. [DOI] [PubMed] [Google Scholar]
  22. Nistér M., Heldin C. H., Wasteson A., Westermark B. A glioma-derived analog to platelet-derived growth factor: demonstration of receptor competing activity and immunological crossreactivity. Proc Natl Acad Sci U S A. 1984 Feb;81(3):926–930. doi: 10.1073/pnas.81.3.926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Paulsson Y., Hammacher A., Heldin C. H., Westermark B. Possible positive autocrine feedback in the prereplicative phase of human fibroblasts. Nature. 1987 Aug 20;328(6132):715–717. doi: 10.1038/328715a0. [DOI] [PubMed] [Google Scholar]
  24. Robbins K. C., Antoniades H. N., Devare S. G., Hunkapiller M. W., Aaronson S. A. Structural and immunological similarities between simian sarcoma virus gene product(s) and human platelet-derived growth factor. Nature. 1983 Oct 13;305(5935):605–608. doi: 10.1038/305605a0. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Rönnstrand L., Beckmann M. P., Faulders B., Ostman A., Ek B., Heldin C. H. Purification of the receptor for platelet-derived growth factor from porcine uterus. J Biol Chem. 1987 Mar 5;262(7):2929–2932. [PubMed] [Google Scholar]
  27. Sejersen T., Betsholtz C., Sjölund M., Heldin C. H., Westermark B., Thyberg J. Rat skeletal myoblasts and arterial smooth muscle cells express the gene for the A chain but not the gene for the B chain (c-sis) of platelet-derived growth factor (PDGF) and produce a PDGF-like protein. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6844–6848. doi: 10.1073/pnas.83.18.6844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stroobant P., Waterfield M. D. Purification and properties of porcine platelet-derived growth factor. EMBO J. 1984 Dec 1;3(12):2963–2967. doi: 10.1002/j.1460-2075.1984.tb02241.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Westermark B., Johnsson A., Paulsson Y., Betsholtz C., Heldin C. H., Herlyn M., Rodeck U., Koprowski H. Human melanoma cell lines of primary and metastatic origin express the genes encoding the chains of platelet-derived growth factor (PDGF) and produce a PDGF-like growth factor. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7197–7200. doi: 10.1073/pnas.83.19.7197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Williams L. T., Tremble P. M., Lavin M. F., Sunday M. E. Platelet-derived growth factor receptors form a high affinity state in membrane preparations. Kinetics and affinity cross-linking studies. J Biol Chem. 1984 Apr 25;259(8):5287–5294. [PubMed] [Google Scholar]
  32. Yarden Y., Escobedo J. A., Kuang W. J., Yang-Feng T. L., Daniel T. O., Tremble P. M., Chen E. Y., Ando M. E., Harkins R. N., Francke U. Structure of the receptor for platelet-derived growth factor helps define a family of closely related growth factor receptors. Nature. 1986 Sep 18;323(6085):226–232. doi: 10.1038/323226a0. [DOI] [PubMed] [Google Scholar]

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